Volume 10 - American Academy of Forensic Sciences

Transcription

AMERICAN ACADEMY OF FORENSIC SCIENCES
410 North 21st Street
PO Box 669
Colorado Springs, CO 80901-0669
Phone: (719) 636-1100
Fax: (719) 636-5245
E-mail: [email protected]
Website: www.aafs.org
PROCEEDINGS
of the American Academy of Forensic Sciences
The Proceedings of the American Academy of Forensic Sciences is an official publication of the American Academy of
Forensic Sciences (AAFS). It is devoted to the publication of the abstracts of technical oral papers and posters presented at the
AAFS annual meeting. These include various branches of the forensic sciences such as pathology, toxicology, physical
anthropology, psychiatry, immunology, odontology, jurisprudence, criminalistics, questioned documents, and engineering.
Similar submissions dealing with forensic oriented aspects of the social sciences are also included.
Please note that some of the abstracts included in the Proceedings deal with topics, results, and/or conclusions which are
controversial. The publication of abstracts does not necessarily imply that the AAFS, its sections, or the individual section program
chairmen/committee has verified or agrees with the studies, results, and/or conclusions of each abstract. During the process of
planning a scientific program, it is impossible to “peer-review” each abstract and presentation to the degree that is accomplished
during manuscript review. Abstracts and presentations are accepted, in part, so that they can be critiqued and reviewed by other
scientists, and so that a forum is available to discuss controversial issues.
The views expressed in this publication are not those of the AAFS. The data and opinions appearing in the published material
were prepared by and are the responsibility of the contributor(s), not of AAFS or its respective employees, employers, officers,
and agents. The AAFS does not supply copies of meeting papers and it is suggested that you write directly to individual authors
to obtain copies of specific papers. Presentation of some abstracts may have been scheduled or canceled after the publication of
this document.
English is the official language of the AAFS and its meetings; neither oral nor written translations will be
provided.
Copyright 2004 by the AAFS. Unless stated otherwise, noncommercial photocopying of editorial material published in this
periodical is permitted by AAFS. Permission to reprint, publish, or otherwise reproduce such material in any form other than
photocopying must be obtained from AAFS.
Printed in the United States of America by Publication Printers, Corp., Denver, CO.
PROCEEDINGS
of the American Academy of Forensic Sciences
Contents
Financial Disclosure Index ...........................................................1
Special Sessions ............................................................................5
Breakfast Seminars .......................................................................8
Luncheon Seminars.....................................................................10
Workshops & Workshorts ...........................................................11
Scientific Sessions
Criminalistics.........................................................................24
Engineering Sciences...........................................................111
General.................................................................................154
Jurisprudence .......................................................................191
Odontology ..........................................................................199
Pathology/Biology ...............................................................221
Physical Anthropology ........................................................268
Psychiatry & Behavioral Science ........................................333
Questioned Documents........................................................340
Toxicology ...........................................................................346
Last Word.............................................................................376
Presenting Author Index ...........................................................382
Key Word Index ........................................................................387
February 2004
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Financial Disclosure
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As a sponsor of continuing education, the American Academy of Forensic Sciences must insure balance, independence, objectivity, and
scientific rigor in all its educational activities. All faculty participating in a sponsoring activity are expected to disclose to the activity
audience any significant financial interest or other relationship: (1) with the manufacturer(s) of any commercial product(s) and/or
provider(s) of commercial services discussed in an educational presentation, and (2) with any commercial supporters of the activity
(such as grants or research support, employee, consultant, major stockholder, member of speaker's bureaus, etc.). The intent of this
disclosure is not to prevent a speaker with a significant financial or other relationship from making a presentation, but rather to provide
listeners with information on which they can make their own judgments. It remains for the audience to determine whether the speaker's
interest or relationships may influence the presentation with regard to the exposition or conclusion.
A
Patricia J. Aagaard, BS - W3
Applied Biosystems
Marie L. Allen, PhD
Applied Biosystems - B141
Pyrosequencing AB - B55; B97
Swedish Government - B55, B97, B141
William P. Allison, JD - SS1
Court TV
José R. Almirall, PhD - B106
Agilent Technologies
CETAC Technologies, Inc.
Perkins Elmer
Robert N. Anderson, PhD, PE - W11
STARK rxp
B
Stan Bajic, PhD
Federal Bureau of Investigation - B95
National Institute of Justice - B165
Elzbieta Bakowska, PhD - B83
National Medical Services, Inc.
John Ballantyne, PhD - W21
ReliaGene Technologies, Inc.
QIAGEN Inc.
Charles E. Barna, BS - W21
QIAGEN Inc.
Suzanne M. Barritt, MS - W3
Applied Biosystems
Michael R. Baylor, PhD - B105
Drug Enforcement Administration
Michael D. Bell, MD - W12
The Divers Alert Network
Duke University Medical Center
Paulina N. Berdos, BS - B18
National Institute of Justice
Robert A. Bever, PhD - W3
Applied Biosystems
Bhanwar Lal Bhootra, MD - G1
Limpopo, South Africa
Department of Health
Robert P. Biancavilla, MPS, JD
Applied Biosystems
Joan M. Bienvenue, MS - B71
Federal Bureau of Investigation
Promega Corp.
QIAGEN Inc..
Tom E. Bodkin, MA - H91
Hamilton County Medical
Examiner Office
Richard S. Brown, MS - C4
MVA, Inc.
Joseph P. Buckley, MS - SS1
Court TV
Bruce Budowle, PhD - W3
Applied Biosystems
Eric Buel, PhD - B30
John M. Butler, PhD
Applied Biosystems - W3
National Institute of Justice - B147
ReliaGene Technologies, Inc. - W21
QIAGEN Inc. - W21
C
Ismail Cakir, PhD - B2; B49
The Council of Forensic Medicine,
Ministry of Justice, Turkey
William B. Campbell, PhD - J5
Accurate Automation Corp.
Theresa A. Caragine, PhD - B26
Titan, Inc.
Eric J. Carita, BA - B112
Brian D. Carrier, MS - D11
The Center for Education and
Research in Information
Assurance (CERIAS)
John M. Carson, DDS - F24; F25
Limbic Systems, Inc.
James L. Caruso, MD - W12
Duke University Medical Center
The Divers Alert Network
Salih Cengiz, PhD - B5; K10
Istanbul University Research Fund
Ranajit Chakraborty, PhD - W21
QIAGEN Inc.
Daniel P. Cheswick, BS - B160
New York City Office of the Chief
Medical Examiner
Promega Corp.
QIAGEN Inc.
Francis A. Chiafari, MS - B23
Amersham Biosciences
Angi M. Christensen, MA, PhD - H104
University of Tennessee
Meihua Chu, PhD - W21
QIAGEN Inc.
L. Scott Chumbley, PhD - D31
Midwest Forensic Research Center
Denise Chung - B146
Michael D. Coble, PhD - B147
Gail A. Cooper, PhD - K30
Cozart Bioscience, Ltd.
David R. Craig, BS - C36
Meta Environmental, Inc.
Thomas A. Crist, PhD - LW5
National Park Service
Cecelia A. Crouse, PhD - SS1
Court TV
Sharon R. Crowley, RN, MN - D55
CooperSurgical, Inc./Leisegang, Inc.
Byron D. Curtis, BS - K33
University of Oklahoma Department
of Pharmaceutical Sciences
D
Darlene Daniels, MS - B137
Tracey C. Dawson, PhD - B134
Applied Biosystems
Virginia Commonwealth University
Forensic Sciences Program
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Peter de Kniijff, PhD - W21
QIAGEN Inc.
Patrick De Smet, PhD - C57
Ghent University
Gary S. Deegear, MD - D10
Biodynamic Research Corp.
Henry J. Dondero, DDS - F11
Microsoft Corporation
E
Brian A. Eckenrode, PhD - C44
Paul D. Emanovsky, MS - H87
Oak Ridge Institute for Science and
Education (ORISE)
David M. Epstein, BS - B96
ARTEL
F
Todd W. Fenton, PhD - H50; H68
Michigan State University
Michael Finnegan, PhD - H35
Forensic Anthropological Consultants
Roger L. Firor, PhD - K12
Agilent Technologies
Constance L. Fisher, PhD - W3
Applied Biosystems
Anna Foror, BS - B83
Paul A. Foxall, PhD
Applied Biosystems - B138
ReliaGene Technologies, Inc. - W21
QIAGEN Inc.- W21
William E. Frank, MS - W21
QIAGEN Inc.
Laurel Freas, BA - H62
National Science Foundation
Linda Frese, MS - B132
New York City Office of the
Chief Medical Examiner
Dean F. Fritch, PhD - K13
Orasure Technologies, Inc.
Tony Frudakis, PhD - D9
DNAPrint™ Genomics, Inc.
Glenn S. Frysinger, PhD - B126
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Norman Gahn, MFS, JD - SS1
Court TV
Alexi Gapeev, PhD - B87
University of Central Florida
Zach Gaskin, BS - D9
DNAPrint™ Genomics, Inc.
Zeno J. Geradts, PhD - C60
Ministry of Justice
Netherlands Forensic Institute
OMECA
Kerstin M. Gleim, BA, BS - B152
Pacific Coast Forensic Science
Institute, Inc.
Bruce A. Goldberger, PhD - W15
Aegis Sciences Corporation
Mirelle I.M. Goos, MSc - C11
Bill J. Gurley, PhD - W15
Leonor Gusmao, PhD - W21
QIAGEN Inc.
H
Ashley Hall, MS - B154
Deborah K. Haller, BS - B16
Applied Biosystems
Armed Forces Institute of
Pathology/DOD
Joy L. Halverson, DVM, MPVM - B150
Orange County Sheriff’s Department
QuestGen Forensics
Douglas R. Hares, PhD - W3
Applied Biosystems
Albert B. Harper, PhD, JD - B84
Amy P. Hart, MD - G25
Revivant
Raymond K. Hart, PhD, JD - C50
Pratt & Whitney
Susan Hastings, MS - B142
Lorraine E. Heath, BSc - B151
Liverpool John Moores University
Douglas P. Heller, PhD, MBA - B102
National Forensic Science
Technology Center
Carol Henderson, JD - E9
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Nicholas P. Herrmann, PhD - H24
Forensic Anthropology Center,
The University of Tennessee
Katie M. Horsman, MS - B28
Frank S. Horvath, PhD - D28
U.S. Department of Defense
Max M. Houck, MA - B117
David G. Howitt, PhD - C31
University of California
Min Huang, PhD - B171
Edwin F. Huffine, MS - B64
Applied Biosystems
Promega, Corp.
Kimberly A. Huston, BS - W21
QIAGEN Inc.
Dixie L. Hybki, MS - B59
Applied Biosystems
I
Alice R. Isenberg, PhD - W3
Applied Biosystems
J
Jon O. Jacobson, PhD, PE - W11
STARK rxp
Richard L. Jantz, PhD - H13; H48
University of Tennessee Faculty
Grant
Susan W. Jones, PhD, MFS - B29
Applied Biosystems
Armed Forces Institute of
Pathology/DOD
Perkin Elmer
Matthew P. Juhascik, MS - K43
Jane S. Juusola, BS - B148
State of Florida
K
Moshe Kam, PhD - J1
TSWG
Steven B. Karch, MD - W15
Melissa S. Kell, BSc - G7
Pharmacia Corp.
Upjohn Diagnostics AB
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Erin H. Kimmerle, MA - H15
University of Tennessee
Richard L. Kingston, PhD - W15
Stephen J. Kish, PhD - K34
Canadian Psychiatric Research
Foundation
Rijad Konjhodzic, BS - B67
Applied Biosystems
Elias J. Kontanis, BS, BA
Forensic Sciences Foundation, Inc. - H30
Sigma Xi Research Grant - H30; H64
The Scientific Research Society - H30
Thomas Krompecher, MD - G60
Swiss National Science Foundation
L
Carll Ladd, PhD - W21
QIAGEN Inc.
Nana Yaa S. Lamouse-Smith, MS - B23
Molecular Staging, Inc.
Robert Large, PhD - B128
M-Scan, Inc.
M-Scan, Ltd.
Stacey A. Lasseter, MSN, RN - D20
Harris County Medical
Examiner's Office
Gilbert Lau - K44
Slim 10
Jumi Lee, PhD - B123
CRAIC Technologies
Soong Deok Lee - B8
Seoul National University Hospital
Research Fund
Patrick Lento, MD - G22
Ethicon, Inc.
Melissa Li, MD - G10
Allergan, Inc.
Dong-Liang Lin, PhD - K4
Institute of Forensic Medicine
Laura L. Liptai, PhD - W11
STARK rxp
Barbara Llewellyn, PhD - W21
QIAGEN Inc.
Jose A. Lorente, MD, PhD - B162
Whatman, Inc.
* Presenting Author
M
N
Pamela L. Marshall, MS - B4
MiraiBio, Inc.
Paul Martin, PhD - J7
CRAIC Technologies
Arni S. Masibay, PhD, MSFS - B145
Promega Corp.
Julie L. Maybruck, BA, BS - B9
Ohio State University Graduate
Student Alumni Research Award
Christopher M. McDermott, MA - H89
U.S. Army Central Identification
Laboratory
Robin L. McDowell, MFS - B23
Richard W. McLay, PhD - W11
STARK rxp
Timothy P. McMahon, PhD - W21
Qigaen
Anne H. McNamee, DDS, MSc - F34
Adobe Systems, Inc.
American Board of Forensic
Odontology
American Society of Forensic
Odontology
Terry W. Melton, PhD - W3
Applied Biosystems
Harry H. Mincer, DDS, PhD - F23
American Board of Forensic
Odontology
John Moalli, ScD - G22
Ethicon, Inc.
Christine Moore, PhD
Aegis Sciences Corporation - W15
U.S. Drug Testing Laboratories - K25
Melody A. Moore, MS - B58
Applied Biosystems
Betsy Moran, PhD - B50
Whatman, Inc.
Mitchell Morrissey, JD - SS1
Court TV
Robert J. Morton, MS - D24
National Center for the Analysis
of Violent Crime
Martina Nilsson - B140
Applied Biosystems
Pyrosequencing AB
Swedish Government
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O
Richard Ofshe, PhD - SS1
Court TV
P
Walther Parson, PhD - W3
Applied Biosystems
Gabor Patonay, PhD - B115
Jeannette M. Perr, BS - B88
National Forensic Science
Technology Center
Joseph L. Peterson, DCrim - B36
Bureau of Justice Statistics
Eric Phillips, BS - B111; K1
Thermo Electron, Corp.
John V. Planz, PhD - W3
Applied Biosystems
Eric G. Pokorak, BS - W3
Applied Biosystems
Deborah Polanskey, BS - W3
Applied Biosystems
Gerry A. Polverento, BS - D17
Michigan Department of Community
Health Grant
Elayne J. Pope, MA
Forensic Sciences Foundation, Inc. B44; H78
FSF Lucas Research Grant - B44
Lawrence A. Presley, MA, MS - B83
Raymond J. Prime, PhD - B75
Government of Ontario
R
Virginia L. Raker, BS - B136
State of Florida
J. Graham Rankin, PhD - B80
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Alan J. Redd, PhD - W21
Qigaen
Elizabeth N. Richards, MS, PhD - G68
Texas Tech University
Rhonda K. Roby, MPH - B72; B135
Applied Biosystems
Reena Roy, MD - W21
Qigaen
Guy N. Rutty, MD, MBBS - B172
KA Scene of Crime Equipment, Ltd.
John H. Ryan, PhD - B170
Myriad Genetic Laboratories, Inc.
S
Maria Dolores Sanchez, MD - D53
International Criminal Investigative
Training Assistance Program
Christine T. Sanders, BA - B27
FUHS/The Chicago Medical School
Leica Microsystems
Karin Scalise - SS1
Court TV
Jason E. Schaff, PhD - K6
Laboratory
Berry Scheck, JD- SS1
Court TV
George J. Schiro, Jr., MS
Applied Biosystems - B13; B31
QIAGEN Inc. - B31
Kenneth G. Schoenly, PhD - G63
California State University
Benjamin R. Schroeder, BS - B53
Mindy E. Setzer, BS - B149
State of Florida
April A. Shea, BA - B83
Jaiprakash G. Shewale, PhD - B11; W21
Qigaen
Kyoung-Jin Shin, DDS, PhD
Human Identification Research
Institute, Yonsei University - F2
Korea Science and Engineering
Foundation - D5
Gary G. Shutler, PhD - B155
Applied Biosystems
Michael E. Sigman, PhD - B86
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Tal Simmons, PhD - H106
Bournemouth University
Sudhir K. Sinha, PhD - W21
QIAGEN Inc.
Dennis E. Slice, PhD - H21
American Museum of Natural History
Smithsonian Institution
James E. Starrs, LLM - W11
STARK rxp
John E.B. Stewart, PhD - W3
Applied Biosystems
Peter R. Stout, PhD - W15
Clay Strange, JD - SS1
Court TV
Aleksandra E. Stryjnik, BSc - B127
Centre of Forensic Sciences
Grand River Products
Anjali R. Swienton, MFS, JD - SS1
Court TV
Christopher K.C. Syn, PhD - B54
Health Sciences Authority
Research Biolabs, Pte Ltd.
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Allan Tereba, PhD - B145
Promega Corp.
Catherine E. Theisen, PhD - W3
Applied Biosystems
Matthew J. Thomas, PhD - D9
DNAPrint Genomics, Inc.
Leigh B. Thorne, MD - B25
College of American Pathologists
Nicole D. Truesdell, BA - H2
Minority Office of Fellowships
and Grants
Smithsonian Institution
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John Vasiliades, PhD - K3
STC Technologies, Inc.
Arpad A. Vass, PhD - D35
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Lauren E. Way - G3
Have-a-Hart
The Clorox Company
Tidbit
Donna E. Webster, BS - K32
Neogen Corp.
Paul J. Wellman, PhD - W15
Carrie M. Whitcomb, MSFS - BS4
Erica Williams, MD - G23
Becton, Dickinson and Company
Verity Software, Inc.
Mary R. Williams, BS - B100
Jeremy P. Wintz, BS - B48
Marshall University
Steven H. Wise - B10
Technical Support Working Group
Z
Nannepaga Y. Zachariah, PhD - D7
Diagnostic Systems Laboratories, Inc.
U
H. Bülent Üner, PhD
Istanbul University - B1; B92
Netherlands Ministry of Justice - B92
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Gerard J.Q. van der Peijl, PhD - B60; B99
Ministry of Justice
Peter Vasilenko, PhD - G70
First Candle (National SIDS Alliance)
* Presenting Author
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Special Sessions
Multidisciplinary Symposium on the
Uses of Forensic Science -The Anatomy
of a Coerced Murder Confession: Can
Post-Conviction Relief Repair the Integrity
of the Criminal Justice System?
Cecelia A. Crouse, PhD, Palm Beach Sheriff's Crime Laboratory,
3228 Gun Club Road, West Palm Beach, FL 33406; Anjali R.
Swienton, MFS, JD, SciLawForensics, Ltd., 25 Walnutwood Court,
Germantown, MD 20874; William P. Allison, JD, 1012 Rio Grande,
Austin, TX 78701; Joseph P. Buckley, MS, John Reid School of
Interrogation, John E. Reid & Associates, 250 South Wacker Drive, Suite
1200, Chicago, IL 60606; Norman Gahn, MFS, JD, Milwaukee County
District Attorney's Office, 821 West State Street, Milwaukee, WI 532331427; Mitchell Morrissey, JD, Denver District Attorney's Office, 2nd
Judicial District, 201 West Colfax Avenue, Department 801, Denver, CO
80202; Richard Ofshe, PhD, University of California, Berkeley,
Department of Sociology, Barrows Hall (MC1980), Berkeley, CA 947201980; Karin Scalise, Texas Department of Public Safety, P.O. Box 4143,
Austin, TX 78765; Barry Scheck, JD, The Innocence Project; Benjamin
N. Cardozo School of Law, 55 5th Avenue, New York, NY 10003; Clay
Strange, JD, 106 East 6th Street, Suite 900, Littlefield Building, Austin,
TX 78701
On January 17, 2001, Christopher Ochoa was released after serving
12 years in prison. He was convicted after he confessed that he and an
accomplice, Richard Danzinger, committed a murder in 1988. Both men
were convicted and sentenced to life in prison. The problem was that they
did not commit the murder. Ochoa later claimed that the confession was a
result of badgering and threats by police during his interrogation.
Authorities acknowledged there was no other evidence against the two
men, neither of whom had been incarcerated before. In one of many twists
in the case, another man, Achim Josef Marino, who is serving life in prison
for other crimes, wrote a series of letters from 1996 - 1998, to public
officials in which he confessed to the killing in detail.
Attorneys for the two innocent men have criticized officials who
received the letters for not aggressively acting on the information. Despite
Marino's letters, authorities did not begin reinvestigating the case until
1999, after Ochoa contacted the Innocence Project at the University of
Wisconsin Law School, and lawyers there began looking into the matter.
DNA tests performed in 2000 ruled out Ochoa and Danzinger as sources of
semen found in the victim, and implicated Marino.
Though this case was unusual in that the actual perpetrator came
forward, biological evidence had been preserved and was still available for
testing. Ochoa was able to get free representation, and the prosecutor's
office cooperated; such is not always the case.
This session will examine some of the factors that contribute to erroneous convictions of innocent persons including the unreliability of eyewitness evidence, and how interrogation tactics taught to law enforcement
play on psychology to cause innocent people to confess. In addition, recommendations for legislative post-conviction reform and the filing of John
Doe warrants to stay the statute of limitations in no-suspect cases will be
discussed. Finally, in instances where an innocent person is exonerated, not
only are law enforcement and prosecutors left with an overturned conviction, but suddenly a case that may have been closed for decades once
again becomes a live investigation. To address this issue, an integrated
approach to instituting effective cold case squads will be considered.
Coerced Confessions, Post-Conviction Relief, Christopher Ochoa
* Presenting Author
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Young Forensic Scientists Forum —
Non-Traditional Careers in Forensic Science
Claire E. Shepard, MS*, Crime Scene Investigator, DeKalb County
Police Department, 3630 Camp Circle, Decatur, GA 30032; Allison M.
Curran, BS*, Florida International University, 9601 SW 142 Avenue,
#818, Miami, FL 33186; Christopher M. Gojcz, BS*, Drug Enforcement
Administration Special Testing and Research Laboratory, 22624 Dulles
Summit Court, Dulles, VA 22066; Amy Neumann, BS*, Paragon
Genetics, 888 Adelaide Street West, Toronto, Ontario M6J 1B8, Canada;
Amy C. Shaver, BS*, Virginia Division of Forensic Sciences, 6600
Northside HS Road, Roanoke, VA 24019; Sheila M. Estacio, BA*, Office
of the Chief Medical Examiner, New York City Office, Department of
Forensic Biology, 520 1st Avenue, New York, NY 10016; Kenneth E.
Melson, JD*, United States Attorney's Office, 2100 Jamieson Avenue,
Alexandria, VA 22314; José R. Almirall, MS, PhD*, Florida International
University, U.P., Miami, FL 33199; Joseph T. Hefner, BS*, University of
Florida, 4100 SW 20th Avenue, Apartment E-6, Gainesville, FL 32607;
Kenneth G. Furton, PhD*, Florida International University, University
Park Campus, DM 445C, Miami, FL 33199; Jeffrey H. Comparin, BS*, ,
Special Testing and Research Laboratory, 22624 Dulles Summit Court,
Dulles, VA 22066; Gordon A. Angus, BS*, Miami-Dade Police
Department, Forensic Computer Lab, 9690 NW 41 Street, Miami,
FL 33199; Kenneth W. Goddard, MS*, National Fish and Wildlife
Forensics Lab, 1490 East Main Street, Ashland, OR 97520; Steven C.
Batterman, PhD*, School of Engineering and Applied Science, School of
Medicine, University of Pennsylvania, 1964 Cardinal Lake Drive, Cherry
Hill, NJ 08003; Jason H. Byrd, PhD*, Office of the Medical Examiner,
1360 Indian Lake Road, Daytona Beach, FL 32126; Howard Cash, BS*,
Gene Codes Forensics, Inc., 775 Technology Drive, Suite 100A, Ann
Arbor, MI 48108; Ronald L. Singer, MS*, Tarrant County Medical
Examiner's Office, 200 Feliks Gwozdz Place, Fort Worth, TX 76104;
Eve Tokumaru, MSFS*, DNA Secology, 6001 Moon Street NE #3821,
Albuquerque, NM 87111; Eric Stauffer, MS*, MME, Forensic Service
Unit, 1039 Industrial Court, Suwanee, GA 30024; Thomas M.
Blackwell, BS*, Northeast Laboratory, 99 Tenth Avenue, Suite 721, New
York, NY 10011; Susan H. Johns, MA*, Illinois State Police Division of
Forensic Sciences, 630 East Washington Street, Springfield, IL 62701
As its role in society evolves, the field of forensic science continues to
grow and incorporate the skills and knowledge of many different experts
and disciplines. This year's special program will bring to the forefront
some emerging forensic science fields, and will consist of presentations by
established members of the forensic science community. Emphasis will be
placed on areas such as computer crimes, wildlife forensics, forensic engineering, entomology, and the role of the medical examiner's office. The
session will also stress the importance of research and method development, and will serve to make emerging forensic scientists aware of the
opportunities available to them. Complete with a discussion panel on
"Realities of Forensic Science," the session will give students and
emerging forensic scientists an open forum atmosphere in which to discuss
the realities of working as a forensic scientist, and the qualifications necessary to pursue this exciting career.
Following the completion of this special session, the participants
should be aware of the diversity of careers in forensic science, should
understand how to compete for the FSF Emerging Forensic Scientist
Award, should recognize the motivation behind and the guidelines for
accreditation of forensic science academic programs, and should be aware
of the realities in the field of forensic science according to young forensic
scientists and crime laboratory directors. A breakfast session and an
evening session will allow participants to mingle with students and
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emerging scientists from across the nation and to establish relationships
with members for mentoring and networking. While at the breakfast
session, participants will also have a chance to learn about resumé building
and are encouraged to bring their own resumés to be critiqued. While at the
evening session participants will be able to view graduate student posters
and to mingle with their peers in a comfortable environment.
This program should appeal to individuals with a strong desire to enter
the field of forensic science as well as those with a few years of experience
within the field and looking to get ahead. The session aims to cover a wide
range of emerging fields through presentations, discussions, and interactions with new and established members of the forensic community. This
program will provide a well-rounded understanding of different areas
within forensic science.
The objectives of this special session are as follows:
• to introduce emerging fields in forensic science;
• to increase awareness of the accreditation process;
• to introduce emerging forensic scientists and prospective forensic
scientists to established forensic scientists through interaction,
discussions, and presentations;
• to provide an understanding of the FSF Emerging Forensic Scientist
Award;
• to provide an opportunity for an open-forum discussion about
realities in forensic science; and,
• to increase involvement within the Young Forensic Scientists Forum.
Careers, Education, Young Forensic Scientists Forum
SD1
Accreditation of Forensic Science Academic
Programs Through the AAFS FEPAC
José R. Almirall, PhD*, Department of Chemistry, International Forensic
Research Institute, Florida International University, Miami, FL 33199;
Max M. Houck, MA*, West Virginia University, 886 Chestnut Ridge Road,
Suite 211, PO Box 6216, Morgantown, WV 26506-6212; Charles G.
Tindall, PhD*, Metropolitan State College, Department of Chemistry,
PO Box 173362, Campus Box 52, Denver, CO 80217-3362; Carl M.
Selavka, PhD*, NIJ Office of Science and Technology, Investigative &
Forensic Science Division, 810 7th Street, NW, Room 7235, Washington,
DC 20531; Jay A. Siegel, PhD*, Michigan State University, School of
Criminal Justice, 506 Baker Hall, East Lansing, MI 48824-1118
After attending this presentation, attendees will understand the
process of accreditation through the AAFS FEPAC mechanism and be able
to participate in the process as an inspector of academic programs.
This presentation will impact the forensic community and/or
humanity by providing a better understanding of the accreditation process
and to encourage academic institutions to participate in the accreditation
process. We also expect to encourage practitioners to invest in the process
by volunteering to act as a site inspector. Forensic Science education will
ultimately benefit from a bona fide accreditation process.
Attendance to this workshop will assist academic institutions who
offer undergraduate and graduate degree programs in forensic science to
prepare for the AAFS accreditation process through the Forensic Science
Education Programs Commission (FEPAC). The workshop will also assist
future site inspectors (academic and practitioners) in preparation for site
inspections of academic programs.
The mission of the FEPAC is to maintain and enhance the quality of
forensic science education through a formal evaluation and recognition of
college level academic programs. The primary function of the committee is
to develop and maintain standards and administer an accreditation program
that recognizes and distinguishes high quality undergraduate and graduate
forensic science programs.
Accreditation of Academic Programs in Forensic Science, FEPAC,
AAFS
6
ES1
Advocacy For the Novice: How to Work
For Forensic Sciences
J.C. Upshaw Downs, MD*, Georgia Bureau of Investigation Medical
Examiner, 925 A Mohawk Drive, Savannah, GA 31419; Barry A.J.
Fisher, MS, MBA*, Scientific Services Bureau, Los Angeles County
Sheriff's Department, 2020 West Beverly Boulevard, Los Angeles, CA
90057-2401; Elizabeth Lavach*, ELS & Associates, 9322 Old Burke
Lake Road, Burke, VA 22015; Joseph P. Polski*, International Assoc for
Identification, 2535 Pilot Knob Road, Suite 117, Mendota Heights, MN
55120-1120
The goal of this special session is to provide a better understanding of
how to effectively lobby budget makers at the local, state, and federal
levels.
Forensic scientists and medical examiners are staunch advocates for
their respective systems. Over the past few years, the overwhelming need
for adequate resources in both systems has led to an attempt at synergy in
advocacy. This effort has had variable results. While the credentials and
skills of those within the nation's forensic community are unparalleled, negligible training has been available for the community as to why and how to
lobby. Systems vary considerably in structure and operation, with the vast
majority of U.S. forensic services provided at the local and state levels.
Recent federal funding streams have also necessitated approaching congressional and senate members to impress upon them the needs of the U.S.
forensic system.
The foundations of our nation's justice system are threatened by the
current inability of forensic labs and medical examiners to process evidence
quickly due to backlogs, the loss of crime scene expertise due to funding
constraints, and the inability to perform certain tests for a myriad of reasons
(such as lack of equipment, lack of training, and lack of expertise). All of
these shortcomings directly relate to one thing - inadequate resources.
When one realizes that state and local labs analyze over 90% of all forensic
cases in the U.S., and that medical examiners analyze 20% of the ~2.5
million U.S. deaths annually, the potential impact of a continued under
funding on the nation's forensic system is of utmost concern.
In 1999, the American Society of Crime Laboratory Directors
(ASCLD) conducted a critical needs survey which concluded that 9,000
more forensic scientists are needed, $1.3 billion is needed for facility modernization and construction, 26% of forensic labs lack basic computer
systems to track evidence, and $285 million is needed in equipment by laboratories. State and local funding continue to be inadequate, and with
recent hard financial times, threaten to get worse before they get better.
Federal monies have been slow to come and often are targeted (earmarked)
during the legislative process.
For example, the federal Crime Lab Improvement Program (CLIP)
which began in fiscal year 2000 has received a total of $95 million in appropriations. This program is intended to establish or improve forensic
analyses by state or local forensic laboratories. To date, $23.4 million has
gone to DNA backlogs, leaving $71.6 million for general forensic laboratory improvement. Of this, $59 million has been earmarked, leaving 13%
($12.6 million) of the total for competitive grants. The earmarked funds
have been used for education, supplies, equipment, technology, facilities,
and contract services. Unfortunately, the $35 million in the fiscal 2002
budget for the Crime Lab Improvement Program (of which 85% is earmarked) did not begin to make a dent in the national forensic infrastructure
needs.
Efforts to secure additional federal monies concluded in 2000 with the
passage and funding of the Paul Coverdell National Forensic Sciences
Improvement Act (NFSIA). This program is intended to provide funding
to crime laboratories and medical examiners' offices based on population
and crime rates. The program permits funding for expenses related to facilities, personnel, computerization, equipment, supplies, accreditation, certification, education, and training in order to improve the quality and timeliness of forensic science or medical examiner services. NFSIA requires a
* Presenting Author
consolidated State plan for forensic science laboratories. Unfortunately,
despite the authorization for hundreds of millions of dollars to date, only
$10 million has actually been appropriated, largely because some see this
initiative as duplicating that of the CLIP.
Part of the difficulty has been the interpretation by federal entities tied
to funding issues that the nation's forensic community was not unified and
not effective in getting the message across to those in charge of federal
funds. Thus, the Consortium of Forensic Science Organizations (CFSO)
was born. The CFSO represents some 11,000 forensic scientists and
medical examiner/coroners. It consists of the ASCLD, the ASCLD/LAB,
the NAME, the IAI, and the AAFS.
This special session will address the history of the efforts to secure
additional funding for the forensic sciences. Through an interactive
approach, techniques that have proven successful or unsuccessful in lobbying will be demonstrated and discussed. The content will cover local,
state, and federal lobbying efforts and effective communication skills.
Public relations skills and concise message driven delivery will be stressed.
The attendee should leave with a better understanding of the ethics of lobbying; consensus building; and who, what, when, where, how, and why to
lobby those public officials who are responsible for agency budgeting.
Lobbying, Advocacy, Funding
ES2
Forensic Scientific Investigations —
Famous Cases Revisited
Application and Limitations of Forensic
Science: Why Experts Sometimes Disagree
A panel of experts will review several highly publicized cases that one
or more of them have been involved with in recent years and will attempt
to point out, and possibly help to explain, the likely basis for such conflicting testimony. The objective of this presentation will not be to convince
the audience of the truthfulness and validity of one theory vs. another, but
rather to edify skeptics and cynics who believe that there can be only one
plausible and acceptable explanation for any alleged homicide scenario.
Forensic scientists need to be frequently reminded that medicine and
some forensic fields are not absolute sciences. Just as there quite often are
marked differences of opinion among various treating physicians in
dealing with clinical matters, so can there be divergent opinions among
forensic pathologists, psychiatrists, toxicologists, anthropologists, entomologists, criminalists, bloodstain and blood spatter experts, document
examiners, forensic engineers, accident reconstructionists, linguists,
accountants, and other forensic subspecialties. Those experts who love to
use the phrase "absolutely, 100% certain'' when proffering testimony
should be admonished not to do so. No civil or criminal court in the United
States requires that degree of certitude, and it is arguably unethical for an
expert to make such a statement.
Such famous cases as Ted Binion, JFK Assassination, Chandra Levy,
Laci Peterson, Michael Peterson, JonBenet Ramsey, O.J. Simpson, and
Jayson Williams are among the cases to be discussed.
The essential background of each of these murder cases will be presented, and the panelists will present their respective analyses, conclusions,
and opinions. Brief comments and succinct questions from the audience
will then be welcomed.
Forensic Scientific Experts, Expert Testimony, Conflicting Opinions
in Famous Cases
Michael M. Baden, MD*, Forensic Sciences Unit, New York State Police,
Building 22, State Campus, Albany, NY 12226; Henry C. Lee, PhD*,
Connecticut Forensic Laboratory, 278 Colony Street, Meriden, CT
06456; Cyril H. Wecht, MD, JD*, Allegheny County, 542 Fourth Avenue,
Pittsburgh, PA 15219a
Attendees will learn about the role and responsibilities of forensic scientists in the criminal justice system and how to handle their interprofessional relationships with opposing experts.
The great majority of homicide cases do not generate conflicting testimony from forensic pathologists and other forensic scientists. A large percentage of these cases do not entail arguable issues such as cause, manner,
mechanism, place and time of death; hence, the roles of the prosecution's
governmentary-related forensic scientists are quite pro forma. In fact,
many of these cases are disposed of by plea-bargaining, and therefore, no
testimony is ever required. Of those cases that do proceed to trial, most
hinge on nonscientific matters, and it is not necessary for the defense to
obtain and utilize independent forensic science experts.
Among the relatively small percentage of murder cases that do culminate in a full-blown trial, those that involve celebrities, physicians, and
other well-known victims or alleged assailants, and some that become a
national and international cause celèbre for inexplicable reasons, forensic
scientific testimony is very often the most highly controversial and critical
component of the entire courtroom drama. Nothing is more titillating,
exciting, and intellectually provocative than a battle between two or more
competent, experienced, and seasoned medical/scientific experts
expressing directly opposing opinions relating to the circumstances of the
victim's death or the alleged assailant's actions in causing that death.
Who to believe? What is the 'truth?' How can equally qualified experts
arrive at mutually exclusive, divergent conclusions and opinions about
such seemingly straightforward scientific issues as cause and manner of
death? Does this kind of litigious-driven dichotomy indicate that one team
of experts has mistakenly interpreted the physical evidence; or too much
emphasis has been given to one particular area of evidence to the exclusion
of other inconsistent or even contradictory facts and circumstances; or
perhaps the intellectual divergence is simply attributable to economic
greed?
* Presenting Author
7
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DALLAS
BS1
Breakfast Seminars
Theatre of the Absurd
Malcolm Stewart-Morris, MD*, Human Performance, 12130 Skyline
Boulevard, Oakland, CA 94619-2423
After attending this presentation, attendees will understand the need
for the restoration of confidence in the justice system and the part
forensic science can play in bringing that about.
humanity by demonstrating that this august body will see itself as a
powerful voice in bringing about the changes in the judiciary which can
restore public confidence and allow the rule of law to approach that of
justice more reliably.
Bearing in mind the theme for the annual meeting, "Truth and
Justice in the Balance," this presentation is designed to prepare the
ground by suggesting that perceptions of the structure underlying those
concepts have undergone cataclysmic change.
From a sadly mishandled election through 9/11 and the aftermath,
including a controversial war which remains unresolved, truth and
justice have not escaped a period in which the constitution itself has tottered.
In viewing a host of troubling issues in domestic and international
affairs, the question posed by Lincoln as to whether government of, by
and for the people can long survive has taken on new urgency which
demands an answer. Forensic science cannot escape harm if its basis in
the rule of law fails to hold the trust of the people.
The author will outline some of the barriers to the counterbalance
forensic science is aiming to provide.
Disillusionment, Reexamination, Restoration
The trial was inconsistent with fundamental due process requirements. The judge was of questionable competence and integrity, the
defense lawyer's behavior made meaningful defense, at best, difficult.
Forensic psychiatric recommendations were violated. The prosecution
was overzealous and the community sprit biased. While in jail Jack
Ruby was a grossly psychotic man but was not given treatment for his
mental illness.
Jack Ruby, Harvey Lee Oswald, John Hinckley
BS3
Just like the movie "Matrix: Reloaded," the production team of
Pitluck, Kenney, and Starrs will focus on movies dealing with Texas
justice and then some.
This year's breakfast will impart the viewers with an understanding
of crime and the law through a wide variety of movie categories,
including westerns, comedies, dramas, and perhaps even some ordinary
home videos. If Texas movies can't rustle up enough controversy to
supply the production team with material, they will revert to their
archives and show how crime, the law, and justice remains the same
from the 1930s through the present.
Crime, Justice, Movies
The Jack Ruby Case — A Miscarriage
of Justice
Emanuel Tanay, MD*, Wayne State University, 2977 Philadelphia Drive,
Ann Arbor, MI 48103
Upon completion of this presentation, attendees will be aware of
the pitfalls of working with lawyers who do not follow the
recommendations of the scientific experts.
On Friday, November 22, 1963, John F. Kennedy, the President of
the United States, was shot in Dallas, Texas, by Lee Harvey Oswald.
Lee Harvey Oswald was killed by Jack Ruby in the basement of the
Dallas Police Department Headquarters. As the psychiatric expert
retained by Jack Ruby's retrial lawyers, the author had considerable
contact with the Warren Commission, including a number of exchanges
with Justice Warren.
Less than twenty years later, John W. Hinckley, Jr., shot President
Ronald Reagan.
The trial lawyer filed a malpractice lawsuit against a psychiatrist
who discharged Hinckley from a psychiatric hospital before the
shooting. The case did not survive the summary judgment motion but it
did provide access to detailed information about John Hinckley's mental
illness.
In the author's opinion the Hinckley verdict was consistent with the
sense of justice and the letter of the law. This does not appear to be true
in the Jack Ruby case.
This presentation will offer historical information that demonstrates
Jack Ruby was a victim of a miscarriage of justice. He was improperly
charged with first-degree murder even though the evidence clearly
showed that it was manslaughter.
8
Movies: Reloaded
Haskell M. Pitluck, JD*, Retired Judge, Illinois Circuit Court, 573 Lake
Avenue, Crystal Lake, IL 60014; Linda B. Kenney, JD*, Law Offices of
Linda B. Kenney, 2 Bridge Avenue, The Galleria, Atrium Building 5, Red
Bank, NJ 07701; James E. Starrs, LLM*, George Washington University,
720 20th Street Northwest, Washington, DC 20052
BS4
BS2
j
2004
Discover the Electronic Trail of Evil:
It’s All About Evidence, Technology,
and Crime!
Carrie M. Whitcomb, MSFS*, National Center for Forensic
Science, University of Central Florida, PO Box 162367, Orlando,
FL 32816-2367
Attendees will be given examples of the technical aspects of computers, the Internet, and the technological foundation for building a case
against electronic crimes and a glimpse at the future.
The Internet and computers, with all their wonderful capabilities,
have long been a place where evil lurks and where latent digital evidence
remains. The technology is changing rapidly and investigators and
forensic scientists must be skilled in tracking and preserving the digital
evidence. Come to breakfast and learn more than you ever wanted to
know about the technology and the possibilities the future holds related
to digital evidence!
Technology, Digital Evidence, Computers
BS5
Death in Beirut
Richard C. Froede, MD*, 3930 N Placita de la Escarpa, Tucson, Z
85750; John J. McDermott, JD*, Hall, Estill, Hardwick, Gable, Golden,
& Nelson, 1120 20th Street Northwest, Washington, DC; Homer R.
Campbell, Jr., DDS*, Office of the Medical Investigator, MSC11 6030,
1 University of New Mexico, Albuquerque, NM 87131-0001
Participants will have an understanding of a historical kidnapping
and forensic investigation and will gain an appreciation of the amount of
time and effort in an investigation of terrorist activity.
* Presenting Author
The previously untold story of the kidnapping and subsequent
deaths of two hostages, William R. Higgins, LTC, United States Marine
Corps, and William F. Buckley, LTC, United States Army, in Beirut,
Lebanon, will now be told. The investigative findings in the deaths in
1985 and 1989 and the discovery as well as recovery of the bodies can
be revealed. The subsequent identification of the bodies and the postmortem findings will be presented. Details of the "alleged hanging" of
Col. Higgins in July 1989 will be discussed. Video presentations will be
shown of the prepared statement by Col. Higgins, the digital identification by the Federal Bureau of Investigation, the evaluation of the
"hanging," and of the story of Col. Buckley’s activity in Lebanon. An
analysis of the two trials in the United States District Court against the
Islamic Republic of Iran, Ministry of Foreign Affairs, will be presented
by the plaintiff's attorney.
Identification, Kidnapping, Recovery
BS6
Deep in the Heart of Texas Justice
James E. Starrs, LLM*, The George Washington University, 720 20th
Street, NW, Washington, DC 20052
After attending this presentation, attendees will be provided with an
overview of the Texas criminal and civil justice systems in extremis. It
will enable the attendee to better appreciate the complexities of programming in the justice system.
humanity by sensitizing the attendee to the nuances of various
troublesome areas in the criminal and civil justice systems of which
he/she is most likely unaware.
Lone Star Justice?
• What do Lance Armstrong and fleas in a courthouse have in
common? They both have found a home in Texas.
• What do Carlton Dotson and James Grigson, MD, (a.k.a.
"Dr. Death") have in common? They both have been featured in
high profile Texas criminal cases.
• What do Juan Corona and Elmer Wayne Henley have in common?
They rank one and two in numbers of serial murders committed by
them with Texan Henley leading by one.
• What do a funeral urn and Darlene Routier have in common?
They both were featured attractions in a Texas murder investigation.
• What do Joe Cannon, a defense lawyer caught sleeping during the
trial of his client Calvin Burdine, and two consenting adults
committing sodomy in private have in common? They both resulted
in notable legal and popular clamor and also originated in Texas.
• What do John E. Wright, Esq., and three time convicted murderer
Johnny Paul Penry have in common? They have both been fighting
for justice in the courts of Texas with Wright seeking to continue his
20 year long defense of Penry.
• What do death row inmate Delma Banks and former FBI Director
William Sessions have in common? They have joined forces in
contesting the Texas capital murder conviction of Banks on Banks’
appeal to the United States Supreme Court for a stay of execution which was granted.
• What do Susan Basso and Jose Betancourt have in common? They
are both losers in the Texas courts, Betancourt having forfeited his
state lottery winnings of $5.5 million and Basso having her cover of
deceit blown by a capital murder conviction.
• What do gypsum wallboard and a cockatoo have in common? They
both were instrumental in separate Texas criminal investigations,
the wallboard in upending drug enforcement in Dallas and the
cockatoo in providing the essential DNA to convict a burglar/
murderer.
• At this point or, hopefully earlier, you have gotten the message. The
point and purpose of this breakfast seminar is to bring together and
* Presenting Author
to focus upon Texas civil, criminal, and courtroom matters that are
larger than life in the expectation that others might learn from these
lessons in Lone Star justice.
Fraud, Forfeiture, Surveillance
BS7
The Quest for Truth in the Emergency Room
Jennifer M. Lafayette, MD*, and Lawrence T. Park, MD, Massachusetts
General Hospital, Warren 605, 33 Fruit Street, Boston, MA 02114;
Felicia A. Smith, MD*, Massachusetts General Hospital, WACC 812,
15 Parkman Street, Boston, MA 02114
This presentation will enhance the ability to evaluate patients with
mental illness and legal issues in the emergency room.
Many patients with mental illness evaluated in the emergency room
have complex legal issues. Some patients arrive while still in police
custody or have outstanding warrants. Other patients may have such significant violent histories that they cannot be hospitalized in a general
psychiatric inpatient unit. Occasionally children are brought to the emergency room with court orders for psychiatric treatment. Overall, legal
status may not be clear. This panel presentation will describe several
cases in which "the quest for truth" led to complex situations in which
disposition was not clear. It will also describe steps for clarifying and
understanding legal issues in the emergency room.
Emergency Room, Psychiatry, Legal
BS8
Tom Krauss Memorial Bite Mark Breakfast:
Evolution of Bite Mark Analysis in North
America From the 20th to the 21st Century
Robert B.J. Dorion, DDS*, 1 Place Ville-Marie, Suite 11238, Montreal,
Quebec H3B 3Y1, Canada
While bitemark recognition in the living is the responsibility of first
responders, emergency room personnel, physicians, pediatricians, dentists,
nurses and social service personnel, the task is relegated to crime scene
investigators, medical examiners, coroners, pathologists and odontologists
in the deceased.
Not surprisingly, the development of a bitemark protocol resulted
from a survey conducted for the odontology section of the AAFS in 1974,
and subsequent ABFO workshops.
Protocols for photography, the collection, the preservation and the
analysis of bitemarks and the respective guidelines and standards were subsequently established by the ABFO.
Past and current methods for evaluating bitemark evidence and
improvements made in comparison techniques in an effort to improve the
scientific basis for bitemark analysis will be discussed.
Contentious issues, prevention and contamination, and the legal
responsibility and liability of the expert witness will be mentioned.
Most of the illustrations used in the keynote address are from the
soon-to-be published textbook "Bitemark Evidence" edited by the speaker.
A team of eminent forensic scientists from the disciplines of forensic odontology, DNA analysis, pathology and jurisprudence present their views in
the book's highly organised thirteen sections. The 30 chapters unravel all
aspects of bitemark evidence from its history, to recognition/diagnosis, to
DNA, to genotypic comparison of oral bacteria, to expert witness liability.
The 21 contributors from four countries present complementary views on
bitemark evidence. The over 700 photographs include 32 color pages illustrating the many facets of bitemark evidence.
Bitemark Evidence, Bitemark History, Bitemark Techniques
9
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DALLAS
L1
Luncheon Seminars
Networking Lunch
Carol Henderson, JD, Stetson University, College of Law, 1401 61st
Street South, Gulf Port, FL 33707; Anjali R. Swienton, MFS, JD,
SciLawForensics, Ltd., 25 Walnutwood Court, Germantown, MD 20874
Take a productive lunch break on Monday to meet new people and to
establish networking relationships. Box lunches will be provided and participants will engage in "speed networking." The hottest trend to hit the
dating scene in years, "speed dating" has been adapted as the latest craze in
networking. Even if you've never engaged in "speed dating," we'll teach
you to speed network - spending a few minutes with each new contact to
exchange information and interests.
The lunch will provide an enjoyable opportunity to network with many
others - the perfect beginning to a week of exciting activities at the AAFS
annual meeting!
Networking, Relationships, Speed Networking
L2
Life in Death’s Acre
Wm. Paul Phillips, JD, PO Box 10, Huntsville, TN 37756; William M.
Bass III, PhD, University of Tennessee, Department of Anthropology,
Knoxville, TN 37996
Dr. William M. Bass reflects on forensic science, violent death, the
"Body Farm," and almost half a century in the company of corpses.
Dr. Bass is a legend in forensic circles. Nearly a quarter-century ago,
Dr. Bass created the world's first (and still the world's only) laboratory
devoted to human decomposition: the University of Tennessee's
Anthropology Research Facility, or ARF—better known as "the Body
Farm." Ever since its inception, this unique facility has produced pioneering scientific research on the rates of human decay under a wide variety
of conditions. It also serves as the world's only setting where human
corpses are regularly used in controlled scientific tests of cutting-edge
forensic technologies such as ground-penetrating radar, artificial "noses"
for sniffing out clandestine graves, and analytical instruments for gauging
time since death on the basis of biochemical "markers" of decomposition.
The Body Farm also provides a unique and realistic training environment
for FBI agents, crime-lab technicians, homicide detectives, and cadaverdog handlers.
Dr. Bass's unique research facility was thrust into the limelight in 1994
when it received title billing in Patricia Cornwell's best-selling fifth novel,
The Body Farm. Besides providing scientific data for Ms. Cornwell's story,
Dr. Bass's facility made a cameo appearance in her novel — as did a character inspired by Dr. Bass himself.
Besides guiding the research at the Body Farm, Dr. Bass has written or
co-authored more than 200 scientific publications, many of them based
either on the facility's research projects or on murder cases and other mysteries he's helped to prosecute or solve. In October 2003, Putnam published
his career memoir, Death's Acre, co-authored with journalist and documentary filmmaker Jon Jefferson.
L3
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2004
Truth is the First Casualty in Conflicts —
The Kosovo Experience
Wm. Paul Phillips, JD, PO Box 10, Huntsville, TN 37756; Helena
Ranta, DDS, PhD, University of Helsinki, Department of Forensic
Medicine, PO Box 40, Helsinki 00014, Finland
The escalation of violence in Kosovo started in early 1998. While the
jurisdiction of the International Criminal Tribunal for the Former
Yugoslavia (ICTY) was not acknowledged by the Milosevic regime,
several states and human rights organizations were considering ways to
send a team of forensic experts to the region to investigate alleged human
rights violations. The European Union Team was finally issued visas and
the Team arrived in Pristina in mid-October 1998.
On January 16, 1999, after the discovery of 45 victims at the village of
Racak, the Team was given a court order to participate in investigations.
After the NATO air strikes in November 1999 and March 2000, the Team
conducted scene investigations at the gullies in the vicinity of Racak and at
the village. The results of these investigations, together with the autopsy
protocols of January 1999, have been filed as evidence in the trial against
Slobodan Milosevic at the ICTY. The speaker was summoned to testify in
the capacity of a 'Chamber Witness' at the trial in March 2003.
In several instances, misleading information was distributed and even
filed as evidence by the accused in relation to Racak. Also, there were
severe shortcomings in the initial investigations at Racak and during the
autopsies at the Institute of Forensic Medicine, University of Pristina.
These include use of paraffin test for gun shot residue (GSR) analysis,
recovery of foreign objects (bullets and bullet fragments), firing distances,
claims of postmortem mutilation, and handling of bodies. After having testified, the speaker will address some of these issues in the light of the
detailed analysis of forensic evidence and DNA profiles.
Kosovo, Chamber Witness, Misleading Information
Body Farm, University of Tennessee, Anthropology Research Facility
10
* Presenting Author
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DALLAS
W1
Workshops & Workshorts
How to Be a Better Expert Witness
Gary R. Brown, BS*, President, RT Environmental Services, 215 West
Church Road, King of Prussia, PA 19406; James S. Smith, PhD*,
President, Trillium, Inc., 28 Graces Drive, Coatesville, PA 19320;
Mary Maloney Huss, BS, JD*, Partner, Wolf, Block, Schorr & SolisCohen, LLP, Wilmington Trust, 1100 North Market Street, Wilmington,
DE 19801; Robert A. Middleburg, PhD*, Forensic Toxicologist,
National Medical Services, 3701 Welsh Road, Willow Grove, PA 19090;
James S. Smith, Jr., PhD*, Oak Creek, Inc., 60 Oak Creek, Buxton, ME
04093-6375
After attending this presentation, attendees will have a better understanding of the responsibilities and legal aspects when called on to be an
expert witness, and understand the techniques that experts use on expert
witness assignments.
Scientists and engineers are often called upon to be an expert witness.
All too frequently, they are not told by other experts, their peers, or lawyers
the key skills necessary to help in a case. Being an expert witness can be a
high point or low point in a professional's career, so learning the key skills
early is key to professional success. Participants in this session will learn:
• How to scope an assignment;
• How to organize work activities and organize facts and data;
• How to prepare focused expert reports; and
• How to prepare for and deliver deposition and trial testimony
A toxic mold case will be used as a "working case" example
throughout the workshop.
Expert Witness, Expert Reports, Trial Preparation
W2
The Role of Atmosphere in Forensic
Investigation: A Closer Look at the
Environment in Forensic Science
Jason H. Byrd, PhD*, Director of Operations, Volusia County Office of
the Medical Examiner, Office of the Medical Examiner, 1360 Idian Lake
Road, Daytona Beach, FL 32124; Ke Chung Kim, PhD*, Penn State
University, Department of Entomology, 501 ASI Building, University
Park, PA 16802-3508; Gregory S. Forbes, PhD*, The Weather Channel,
300 Interstate North Parkway, Atlanta, GA 30339; Paul Knight, MS*,
State Climatologist for Pennsylvania, affiliated with Penn State
University, PSU 608A Walker Building, University Park, PA 16802-5014;
Joseph P. Sobel, PhD*, Accu-Weather, Inc., 385 Science Park Road, State
College, PA 16803
After attending this presentation, the participant will gain information
about the variety of atmospheric data sources, knowledge concerning the
role of climatic conditions in cases, understanding of the procedures used
in reconstruction of weather conditions for the investigation, and awareness
of expertise in this aspect of forensic science.
The Role of Atmosphere in Forensic Investigation: A Closer Look at
the Environment in Forensic Science Crime and crime scenes are potentially influenced by atmospheric and environmental conditions. During the
forensic investigation of events that occur outdoors, the discovery phase
should consider the weather conditions at the time of the incident. Data
from the atmospheric sciences has changed significantly during the past
decade and knowledge of its availability, utility, and potential limitations
can assist in several parts of a case.
* Presenting Author
j
2004
An overview of the tools and their application by forensic climatologists will be followed by the presentation of several case studies where
reconstruction of atmospheric conditions played a crucial part of both civil
and criminal investigations.
Examples of the use of this science will be shown in weather-related
disasters, such as airplane accidents, tornados, hailstorms, damaging windstorms, and floods where the weather phenomena causing the damage is
very small scale. Because of this, they miss the relatively sparse observing
stations, and their conventional data does not represent the wind speeds and
other conditions at the time and location of the incident. This presentation
will describe the use of radar and other tools that can be used in cases of
aircraft icing, microbursts, tornados, and floods, in which the speaker was
the expert witness.
There will also be a presentation illustrating the use of climatology in
accident related incidences where there had been a discrepancy about the
road design and its safety during hazardous weather conditions, as well as
a presentation of how weather conditions influence cases involving
forensic entomology.
Atmospheric Conditions, Climatology, Environmental Impacts
W3
Forensic Human Mitochondrial
DNA Analysis
Douglas R. Hares, PhD, DNA Unit II, FBI Laboratory, 2501
Investigation Parkway, Quantico, VA 22135; John E.B. Stewart, PhD*
and Patricia J. Aagaard, BS*, National Missing Person DNA Database,
FBI Laboratory, DNA Unit II, 2501 Investigation Parkway, Quantico, VA
22135; Suzanne M. Barritt, MS*, Armed Forces DNA Identification
Laboratory, 1413 Research Boulevard, Rockville, MD 20850; Robert A.
Bever, PhD*, The Bode Technology Group, 7364 Steel Mill Drive,
Springfield, VA 22150; Robert P. Biancavilla, MPS, JD*, Nassau County
District Attorney's Office, 262 Old County Road, Mineola, NY 11501;
Bruce Budowle, PhD*, FBI Laboratory, 2501 Investigation Parkway,
Quantico, VA 22135; John M. Butler, PhD*, National Institute of
Standards and Technology, 100 Bureau Drive, MS 8311, Building 227,
Room A243, Gaithersburg, MD 20899-8311; Connie L. Fisher, PhD*
and Alice R. Isenberg, PhD*, FBI Laboratory, DNA Unit II, 2501
Investigation Parkway, Quantico, VA 22135; Terry W. Melton, PhD*,
Mitotyping Technologies, LLC, 1981 Pine Hall Drive, State College,
PA 16801; Walther Parson, PhD*, Institute of Legal Medicine,
University Innsbruck, Muellerstr. 44/III, A-6020 Innsbruck, Austria;
John V. Planz, PhD*, DNA Identity Laboratory, University of North
Texas, HSC, 3500 Camp Bowie Boulevard, Fort Worth, TX 76107;
Eric G. Pokorak, BS*, Deborah Polanskey, BS*, and Catherine E.
Thiesen, PhD*, BS, FBI Laboratory - DNA Unit II, National Missing
Person DNA Database, 2501 Investigation Parkway, Quantico,
VA 22135
research currently being conducted in the field, what technologies are on
the horizon, legal issues faced by the forensic mtDNA community, and
issues encountered in forensic mtDNA casework. Participants should also
have a complete understanding of SWGDAM's Guidelines for
Mitochondrial DNA Nucleotide Sequence Interpretation and the databasing
efforts taking place in the mtDNA community, including missing persons
and population data.
11
humanity by reviewing the current issues in mitochondrial DNA analysis as
well as exploring future technologies in the field.
Over the past decade, mitochondrial DNA (mtDNA) analysis has
become more widely used in the forensic community. Mitochondrial DNA
analyses are particularly important in cases where nuclear DNA analyses
are not possible due to insufficient sample quantity and/(r quality, or lack
of appropriate reference samples. While mtDNA analyses do not provide
the discrimination potential of nuclear DNA tests, mtDNA sequence is
often the only information which can be gathered from a piece of evidence.
This workshop will have three main objectives. The first objective
will be to discuss issues in traditional mtDNA casework. Topics will
include a discussion of The Scientific Working Group on DNA Analysis
Methods (SWGDAM) published document containing guidelines for the
interpretation of mtDNA sequence data, casework examples, heteroplasmy,
the mtDNA population database maintained by the FBI Laboratory, the
Combined DNA Indexing System Missing Person component (CODISmp)
and the FBI's Regional Mitochondrial DNA Program. The second objective
will be to discuss current research in the field of mtDNA. Topics will
include single nucleotide polymorphisms (SNP's), automation, mixtures,
phylogenetics, and possible future technologies. The third objective will be
to discuss legal issues encountered in the forensic mtDNA arena. Panel discussions will allow attendees to address their specific questions and concerns to the speakers after both the morning and afternoon sessions.
mtDNA, Missing Persons, Mitochondrial DNA
W4
Forensic Science, Evidence Based Medicine,
and the “Shaken Baby Syndrome”
Patrick E. Lantz, MD*, Wake Forest University Health Sciences,
Department of Pathology, Medical Center Boulevard, WInstom-Salem,
NC 27157-1072; Thomas L. Bohan, PhD, JD*, MTC Forensics, 371 Fore
Street, Portland, ME 04101; G.G.W. Adams, BSc, MBChB*, Mooresfields
Eye Hospital, City Road, London, EC1V 2PD, United Kingdom; Faris A.
Bandak, PhD*, Department of Neurology, F. Edward Hébert School of
Medicine, Uniformed Services University of the Health Sciences, PO Box
60060, POTOMAC, MD 20854; Patrick D. Barnes, MD*, Lucile Satter
Packard Children's Hospital, Stanford University Medical Center, 725
Welch Road, Palo Alto, CA 94304; Mark Donohoe, MBBS*, 129A
Raglan Street, PO Box 328, Mosman, NSW 2088, Australia; Jan E.
Leestma, MD*, The Children's Memorial Hospital, 1440 North Kingsbury
Street, Suite 210, Chicago, IL 60622; Colin Smith, Neuropathology Unit,
Department of Pathology, The University of Edinburgh, Western General
Hospital, Crewe Road, Edinburgh, EH4 2XU, United Kingdom; Kirk
Thibault, PhD* and Lawrence E. Thibault, ScD*, Biomechanics, Inc.,
Philadelphia Naval Shipyard, Quarters M-2, 4601 South Broad Street,
Philadelphia, PA 19112
After attending this presentation, attendees will from an evidence
based medicine perspective, be able to discuss the quality of evidence limitations regarding the existing literature on 'Shaken Baby Syndrome'
including retinal hemorrhages/ocular manifestations, radiographic findings,
traumatic unconsciousness, criteria for case definition, and methodological
flaws/selection bias in study design; and be able to summarize the biomechanics in age-related traumatic brain injury including causal mechanisms,
scaling of tolerances for skull failure and brain injuries in adults and
children, and limitations of diagnostic assumptions without biomechanical
considerations.
12
More than three decades have passed since John Caffey's publication
"On the Theory and Practice of Shaking Infants." Since then the 'Shaken
Baby Syndrome' (SBS) has received extensive clinical attention as well as
national and international publicity. Because of a number of highly publicized court cases, pathologists, pediatricians, ophthalmologists, radiologists, neurosurgeons and biomechanical engineers are being critically scrutinized about their opinions on this subject. No one can dispute that the protection of innocent children is beneficial for society; however, this must be
grounded in objective, unbiased and reproducible scientific facts thereby
permitting a balance between truth and justice. If not, others rights are
endangered, the wrongfully accused.
Inertial loading with abrupt angular acceleration/deceleration during
shaking has been postulated to cause cerebral shear strain deformation, tissue
disruption at interfaces and diffuse (traumatic) axonal injury. Such a mechanism seems inherently logical in susceptible infants and young children who
would be inclined to this type of injury by relatively weak neck muscles, a
proportionally large head and vulnerability of an immature brain.
Traditionally, this shaking mechanism hypothesis has assumed to explain the
primary injury patterns (subdural hematoma, retinal hemorrhages and diffuse
axonal injury) and secondary injury patterns (cerebral edema,
hypoxia/ischemia and herniation) stated to be characteristic if not virtually
pathognomonic for SBS.Presumably causing immediate symptoms without
a lucid interval these serious and potentially life-threatening injuries are often
readily demonstrated by neuroimaging, funduscopic examination and neuropathologic procedures. According to a number of medical articles, similar
accidental cerebral/ocular injuries can only occur with massive forces associated with motor vehicular collisions or a fall from at least a two-story
building. Also, an infant or child with a prior head injury who has changing
symptoms would presumably indicate a newly inflicted injury and not an
intracranial rebleed. Other authors have stated that "when extensive retinal
hemorrhages accompanied by perimacular folds and retinoschisis are found
in association with intracranial hemorrhage or other evidence of a traumatic
brain injury in an infant, a shaking injury can be diagnosed regardless of other
circumstances." With this reasoning, the last caregiver attending the injured
infant/young child is automatically considered guilty of causing an abusive
head injury, especially if unwitnessed since shaking is never considered accidental. This dilemma has been exacerbated by the absence of a consistent,
reliable case definition for the diagnosis of SBS and the lack of evidence base
within the SBS literature. Much of the current medical literature on SBS consists of case reports, non-comparative case series, unsystematic reviews and
position papers. Inclusion criteria in a number of case reports and case series
include vague diagnostic categories such as 'presumed' child abuse. Case
control studies often exhibit selection bias in methodological design and circular reasoning. The only experimental biomechanical study of SBS measured forces from shaking versus impact in a doll model. Only forces
observed with impact exceeded injury thresholds that would explain the
pathological spectrum of the intracranial injuries. Although subsequent
reports have corroborated that intracranial injuries are associated with impact,
they also offered other evidence that shaking alone is sufficient to cause
serious injury and/or death in infants and young children.
The first part of this workshop critically reviews the scientific literature on SBS including retinal hemorrhages/ocular manifestations, radiographic findings, and traumatic unconsciousness. The subsequent sessions
review the clinical and biomechanical literature about the basic mechanisms of adult and pediatric head injury and address the question whether
shaking alone is sufficient to explain the neuropathologic and ophthalmologic findings. Questions and comments from the audience are encouraged
following each presentation and a panel discussion directed by questions
from the audience concludes this informative workshop.
Evidence Based Medicine, Shaken Baby Syndrome, Traumatic Brain
Injury
* Presenting Author
W5
Tryptamines and Other Psychotropic
(Mind Altering) Substances: Analysis,
Toxicology, and Pharmacology
Peter T. Ausili, BS, MSBA*, Drug Enforcement Administration, North
Central Laboratory, 536 South Clark Street, Suite 800, Chicago, IL
60605; Adam Negrusz, PhD*, Department of Pharmacodynamics,
University of Illinois, 833 South Wood Street, Chicago, IL 60612; Terry
A. Dal Cason, MS*, Drug Enforcement Administration, North Central
Laboratory, 536 South Clark Street, Suite 800, Chicago, IL 60605; A.
Karl Larsen, Jr., PhD*, Illinois State Police, Division of Forensic
Services, 1941 Roosevelt Road, Chicago, IL 60608-1229; R. Francis
Schlemmer, PhD*, Department of Biopharmaceutical Sciences, College
of Pharmacy, University of Illinois at Chicago, 833 South Wood Street,
Chicago, IL 60612; Srihari R. Tella, PhD*, Drug Enforcement
Administration, Drug and Chemical Evaluation Section, Office of
Diversion Control, 2401 Jefferson Davis Highway, Alexandria,
VA 22301
After attending this presentation, workshop attendees will have an
awareness of the current drug abuse trends regarding new and unusual psychotropic drugs. Analytical data, pharmacological effects, and toxicological
consequences will be emphasized.
In the last few years there has been a significant increase in the use,
abuse and availability of new and different psychotropic drugs. These
substances, commonly referred to as "Club Drugs" are not limited to
LSD, DMT, MDA, MDMA (ecstasy), methoxyamphetamines (PMA,
PMMA), Ketamine and GHB. Other new substances being encountered/abused more frequently include designer Tryptamines such as
alpha-Methyltryptamine (AMT), 5-Methoxy-N,N diisopropyltryptamine
(5-MeODIPT); Phenethylamines such as 2C-T-7, 2C-B (Nexus) and its'
analogues 2C-C or 2C-I; and, Piperazines such as Benzylpiperazine
(BZP) and Trifluromethylpiperazine (TFMPP). An example of an
unusual substance that will be discussed is 'Ayahuasca', a South
American herbal extract that contains Dimethyltryptamine (DMT) in
combination with Harmine and Harmaline. Also presented will be polydrug mixtures such as MDA/MDMA/Ketamine/Methamphetamine and
examples of unusual synthesis reaction by-products.
This presentation will examine pharmacological effects, toxicity and
current evaluation and control issues. Analytical reference data will be provided to all workshop attendees. This workshop is designed for practitioners in medicine, forensic sciences (medical examiners/pathologists,
chemists, behavioral psychologists, and toxicologists) and law enforcement
personnel.
Psychotropic Substances (Club Drugs), Toxicology and
Pharmacology, Evaluation and Control
W6
Learning to Appreciate and
Respect Differences
Carol E. Henderson, JD*, Shepard Broad Law Center, Nova Southeastern
University, 3305 College Avenue, Ft. Lauderdale, FL 33314; Anjali R.
Sweinton, MFS, JD*, SciLaw Forensics, Ltd., 25 Walnutwood Court,
Germantown, MD 20874; Carlton-Jane Beck, MS, Lake County Sheriff's
Office, Crime Scene Investigations, 360 West Ruby Street, Tavares, FL
32778; Ashraf Mozayani, PhD, PharmD, Harris County Medical
Examiner Office, 1885 Old Spanish Trail, Houston, TX 77054; Joye
Carter, MD*, 3390 Ozark Street, Houston, TX 77021-1135; Mary Fran
Ernst, BLS*, Saint Louis University School of Medicine, Division of
Forensic Pathology and Education, 6039 Helen Avenue, St. Louis, MO
63134; Susan Hart Johns, MA*, Illinois State Police, Division of Forensic
Services, 630 East Washington Street, Springfield, IL 62701; Patricia T.
Kelly*, 8029 Wilson Terrace, Springfield, IL 62707; Yona Rozen, JD*,
Gillespie, Rozen & Watsky, 3402 Oak Grove Avenue, Suite 200, Dallas,
* Presenting Author
TX 75204, Teri Stockholm, PhD*, The Wise Woman Group, 1700SE 15th
Street, Suite 309, Ft. Lauderdale, FL 33316
By attending this presentation, attendees will be taught skills and
strategies to successfully integrate the varied components necessary for a
successful career in forensic science.
The demographics within forensic science are changing. Many of the
positions in the crime laboratories and graduate schools are now being
sought and held by women. As a result, it is important that the profession
examine the differences that the demographic shift brings to forensic
science. This workshop will feature speakers and activities addressing the
skills and knowledge needed to be successful in this diverse environment.
Topics include: mentoring and networking; communication skills; legal
issues in the workplace (e.g., sexism, ageism, harassment); personal
choices and balancing responsibilities; and gender stereotypes, values and
standards. The workshop will include discussions on surviving politics in
the workplace and overcoming adversity. Participants will be taught skills
and strategies to successfully integrate the varied components necessary for
a successful career in forensic science. Registration for the workshop
includes a luncheon at The Women's Museum in Dallas, a Smithsonian
affiliate, as well as a tour of the museum.
Mentoring, Networking, Communication
W7
Drug Enforcement Administration
Forensic Drug Chemist Workshop
Rhesa G. Gilliland, MS*, Drug Enforcement Administration, Office of
Forensic Sciences, 2401 Jefferson Davis Highway, Alexandria, VA 22301;
Darrell L. Davis, BS*, Laboratory Director, Drug Enforcement
Administration South Central Laboratory, 10150 East Technology
Boulevard, Dallas, TX 75220-4377; Sanford Angelos, MS, MEd*, Drug
Enforcement Administration, 536 South Clark, Suite 800, Chicago, IL
60605-1526; John S. Chappell, PhD*, Drug Enforcement Administration
Western Laboratory, 390 Main Street, Room 700, San Francisco, CA
94105; Tamara D. Keller, BS*, Drug Enforcement Administration, 10150
East Technology Boulevard, South Central Laboratory, Dallas, TX;
Walter R. Rodriguez, MS*, Drug Enforcement Administration, 5202 NW
84th Avenue, Miami, FL 33166; Jerry A. Walker, BS*, Drug Enforcement
Administration Mid-Atlantic Laboratory, 1440 McCormick Drive, Largo,
MD 20774-5313
After attending this presentation, attendees will be able to utilize information and material provided to improve current drug analysis procedures
within their laboratory, to select appropriate instrumental techniques for
controlled substance identification and quantitation, and to receive an
overview of the DEA approach to controlled substance analysis.
humanity by providing an opportunity for crime laboratories to interact
with DEA forensic chemists concerning the challenges of controlled substance identification techniques.
The objective of this workshop is to present a synopsis of the methods
currently being used in DEA laboratories to identify suspected controlled
substances and other materials encountered in evidentiary drug analysis.
DEA has been training individuals in controlled substance analysis
since the early 1970s. Over 5,000 students have been trained either during
state and local seminars or within the DEA laboratory system. These
students have been trained in the use of the most up to date analytical techniques in forensic drug chemistry. DEA forensic chemists form the core
group responsible for presentations during the seminars or within the DEA
field laboratories.
This workshop will be an abbreviation of the DEA Forensic Chemists
Training Program. The presentations will include information which is currently disseminated in the state and local seminars and by each DEA
training officer within the field laboratories. The students will receive infor13
mation on screening techniques to include color tests, microcrystalline
tests, and thin layer chromatography; basic quantitative analysis including
the use of gas chromatography, high performance liquid chromatography
(HPLC), and capillary electrophoresis (CE); and confirmatory instrumental
techniques to include Fourier Transform Infrared Spectrophotometry and
gas chromatography/mass spectroscopy (GC/MS).
It would be impractical to attempt to include all facets of these techniques in a one-day presentation. However, the presentations will cover as
much information as possible during the course of the seven hours of
instruction. At a minimum, the following topics will be included:
• Advantages in the use of one type of analytical technique over
another technique.
• Data evaluations problems which analysts and technical reviewers
must consider prior to reaching conclusions
• Derivatization techniques and instrumental methods used to identify
isomers
• Recommendations for quantitative methodology
The session will conclude with a 90-minute discussion hosted by the
DEA Forensic Chemist instructors. Participants will be invited to ask questions, comment on the information which has been provided, and suggest
solutions for problem solving techniques.
Controlled Substance, Drug Analysis, Instrumental Techniques
W8
Setting Up a Digital Evidence Unit
and Preparing for Accreditation
Carrie M. Whitcomb, MSFS*, SWGDE, PO Box 162367, Orlando, FL
32816-2367; Robert P. Bianchi, BS*, RCFL National Program Office,
5502 Chester Mill Court, Fairfax, VA 22030; Don Lewis*, Lakewood
Police Department, Lakewood, CO 80226; Michael J. Phelan, MA*,
Drug Enforcement Administration Computer Forensics Lab, 7704 Old
Springhouse Road, McLean, VA 22102-3494; Mark Pollitt, MS*, Digital
Evidence Professional Services, 8509 Nicole Court, Ellicott City, MD
21043; Dara Sewell*, Testing and Validation, Unit Chief, FBI CART, 935
Pennsylvania Avenue, Washington, DC 20535
After attending this presentation, attendees should be able to understand the process and criteria for developing a digital evidence unit.
Additionally they will learn about the accreditation process and will have a
fundamental understanding of the basic principles required to develop a
training manual, a quality manual and standard operating procedures
manual for digital evidence.
This workshop will present an overview of establishing a digital
evidence unit and developing quality programs that assist in meeting
accreditation criteria for the unit.
Members of The Scientific Working Group on Digital Evidence
(SWGDE) will present a workshop that describes the process from
deciding to implement a digital evidence unit to its accreditation. The panel
of experts will present the history of digital evidence, performing a needs
assessment, and developing an implementation plan, designing the infrastructure for a quality digital evidence unit capable of achieving accreditation by The American Society of Crime Laboratory Directors /Laboratory
Accreditation Board (ASCLD/LAB).
The American Society of Crime Laboratory Directors/Laboratory
Accreditation Board (ASCLD/LAB) in 2003 approved Digital Evidence as
a discipline in Crime Laboratories. SWGDE has defined digital evidence as
"information of probative value that is stored or transmitted in a binary
form." Sub-disciplines within digital evidence are:
• Computer Forensics
• Digital Audio Forensics
• Digital Images/Video Forensics
SWGDE has developed best practices and examples for digital
evidence quality manuals, training manuals, and SOP manuals to assist
laboratories develop a quality program. These documents will be intro14
duced to the forensic community at this workshop and will assist attendees
in preparing for digital evidence for accreditation.
As a matter of clarification, digital pictures of a crime scene are not
considered to be digital evidence, unless the digital picture itself is the evidence, for example a digital picture or digital video of child pornography.
Digital Evidence, Accreditation, Quality Programs
W9
Serial Homicide, Myths, Legends,
and Facts
Robert J. Morton, BA, MS*, FBI, National Center for the Analysis
of Violent Crime, FBI Academy, Quantico, VA 22135; Mary CollinsMorton, BS*, FBI Washington Field Office, 601 4th Street,
Northwest, Washington, DC 20535; James J. McNamara, MS*,
Alan C. Brantley, MA*, and James O. Beasley, MPA*, FBI, National
Center for the Analysis of Violent Crime, FBI Academy, Quantico,
VA 22135
After attending this presentation, attendees will have a greater understanding of the "truth" about serial homicide, to include the scope of serial
murder, motivations, methods of operation, victim selection, body disposal,
forensic techniques, and cooperative investigative strategies useful to
successful case resolution.
humanity by exposing participants to empirical and case data concerning
serial murder and the unique behavioral, forensic, and investigative issues
facing professionals participating in serial murder investigations.
The purpose of this workshop is to provide investigators and medicolegal professionals with an understanding of serial murderers, their
motives, methods of operation, victim selection, and body disposal scenarios highlighted through case examples. The focus of the workshop is on
the practical issues involved in investigating and analyzing the actions of a
serial murderer, the benefits of input from different disciplines and the need
for cooperation between professionals.
This workshop is targeted at providing investigators and medico-legal
practitioners with a broad base of knowledge concerning serial murder, as
well as a thorough understanding of the nature of serial offenders.
Workshop will include a discussion of serial murder and its parameters,
motivations of serial offenders, forensic issues and investigative issues.
Workshop discussions will be augmented by numerous case examples,
including several "high profile" cases.
Serial homicide has long been an issue that generates much attention,
from law enforcement, mental health practitioners, medico-legal professionals, and the media. There are a plethora of opinions. Law enforcement
regard serial homicide as a painful anomaly, mental health practitioners
look for causes, medico-legal professionals struggle with the results and the
media inflates serial homicide into an epidemic. It's no surprise that many
myths and legends abound concerning serial killers.
The FBI's National Center for the Analysis of Violent Crime
(NCAVC) is routinely consulted by federal, state, and local authorities in a
variety of cases of violent crimes, especially bizarre or serial homicides.
The NCAVC has had extensive experience in assisting federal, state, and
local law enforcement agencies in the investigation of serial homicides, and
has reviewed hundreds of serial homicide cases for research purposes.
Currently, the NCAVC is engaged in several research projects on serial
offenders, including interviews of incarcerated serial offenders. The
material presented in this workshop is based upon actual case experience,
ongoing research, and current interviews with serial offenders.
Upon completion of this workshop, participants can expect to have a
greater understanding of the "truth" about serial homicide, to include, the
scope of serial murder, serial offenders and motivation, methods of operation, victim selection, body disposal, forensic techniques and collection
strategies pertinent to serial homicide, and cooperative investigative
strategies useful to successful case resolution.
Serial Homicide, Serial Murderer, Victimology
* Presenting Author
W10
Forensic Histopathology
W11
Engineering Evidence and Lay Testimony
Paul Fornes, MD, PhD*, Department of Pathology, European Hospital
Georges Pompidou, 20, Rue Leblanc, Paris, 75015, France; Steven
Karch, MD*, PO Box 5139, Berkeley, CA 94705-0139; Bernd
Brinkman, MD, PhD*, International Journal of Legal Medicine,
Rontgenstrausse 23, Munster, D-48149, Germany; Chris M. Milroy MD*,
University of Sheffield, Sheffield S3 7ES, United Kingdom; Helen L.
Whitwell, MD*, Department of Forensic Pathology, University of
Sheffield, Medico-Legal Centre, Watery Street, Sheffield, United Kingdom
Richard W. McLay, PhD, PE*, Professor of Biomedical Engineering,
University of Iowa, 1231 Hamilton Court, Iowa City, IA 52245; Robert N.
Anderson, PhD, PE*, RNA Consulting, Inc., 27820 Saddle Court, Los
Altos Hills, CA 94022; Jon O. Jacobson, PhD, PE*, Jacobson Engineers,
5220 Roosevelt Way, NE, Seattle, WA 98105; Laura L. Liptai, PhD*,
Biomedical Forensics, 1660 School Street, Suite 103, Moraga, CA 94556;
James E. Starrs, LLM*, Professor of Law & Forensics, George
Washington University, 720 20th Street, NW, Washington, DC 20052
After attending this presentation, pathologists should be able to recognize and interpret main histologic lesions in forensic pathology. Nonpathologists should be able to understand and/or criticize a pathology
report.
Surgical/general histopathology textbooks deal with diseases encountered in hospitals, but forensic issues are never addressed. On the other
hand, in forensic textbooks, gross pathology, but rarely histopathology, is
found. Forensic histopathology is also rarely taught in medical schools.
Consequently, it may be difficult for a pathologist to get sufficient
knowledge in forensic histopathology. In this workshop, main histologic
lesions encountered in forensic practice, in different fields, will be
examined.
Natural deaths account for half of autopsies in forensic practice.
Among those, cardiovascular/sudden deaths are by far the leading causes
of death. Examination of the heart is therefore of the greatest importance.
Although some cardiac diseases may be diagnosed at gross examination,
histology may prove useful in certain cases, such as myocarditis, arrhythmogenic right ventricular cardiomyopathies, conduction tissue lesions. The
attendee will be able to examine a heart. Guidelines in sampling the heart,
including conduction tissue will be provided. Main histologic lesions
encountered in forensic practice will be displayed.
Toxic-related deaths are also frequent in forensic practice. In many
cases, toxicology does not allow a clear-cut conclusion, because lethal substance blood levels are usually difficult to assess. Complete histology of
organs should be systematically performed, because not only acute but also
chronic lesions may have contributed to the fatal outcome. These issues
will be examined, and correlation between toxicology and pathology will
be discussed.
Gross examination of the formalin fixed brain is essential, but in many
cases, histology provides useful information. Vascular malformations, non
visible with the naked eye, may sometimes explain parenchymal hemorrhages, which can be misdiagnosed as traumatic. On the other hand, a
trauma may contribute to the rupture of a vascular malformation. Neuronal
necrosis, degenerative diseases, etc., require histologic examination.
Immunohistochemistry is essential in diagnosing diffuse axonal injury,
which is an important issue in forensic practice. The shaken baby syndrome
requires meticulous handling of the brain. Eye histology is essential in this
field. Guidelines in sampling the brain, performing immunohistochemistry
will be provided. Histologic lesions of forensic interest will be displayed.
Diagnoses of mechanical asphyxia and drowning are difficult tasks in
forensic practice. Although histologic lesions are usually not specific, histology of all organs always contributes to the diagnosis in addition with
other findings (autopsy, toxicology, biochemistry, diatoms). Interpretation
of histologic lesions will be discussed.
Dating wounds is a main concern in forensic pathology. Specific
stains and immunohistochemistry can be applied. Up-to-date techniques
will be examined.
Other issues will be finally illustrated by didactic cases. The attendee
will be able to ask questions related to other issues, such as artefacts,
autolysis, etc.
After attending this presentation, attendees will understand
engineering evidence methods and evaluate lay engineering testimony.
This program studies the precedents in the law for the application of
engineering to accident reconstruction. Examples of lay engineering
testimony are shown and discussed relative to the rules of evidence. A panel
discussion reviews methods for combatting questionable lay engineering
testimony. Presentations include:
• Precedents in accident reconstruction law
(The history of lay engineering testimony)
• A criminal case with lay engineering testimony
(A defendant is convicted with improper momentum analyses and
astounding human factors data)
• Precedents in the law of biomechanics testimony
(Examples of questionable lay biomechanics testimony)
• Failure analysis methods in accident reconstruction
(Examples of questionable lay metallurgical and materials science
testimony)
• An example from the law
(The Frank Olson case will be presented)
The panel will tie all of the presentations together by discussing
several practical methods for combating questionable lay engineering
testimony:
• The use of a deposition for sending a layman through an engineering
oral exam, followed by a Daubert challenge;
• The use of the Racketeer Influenced Corrupt Organization (RICO)
Statutes to pursue the municipalities that make use of questionable lay
engineering testimony to obtain criminal convictions;
• The use of the ethics statutes, following a technical deposition of the
expert, showing that the attorney has concealed evidence or otherwise
violated a discovery order, having made misrepresentations to the
court and clearly knows or should have known otherwise;
• Reporting the lay engineering testimony to the appropriate state
licensing board with the objective of both criminal and civil actions
for practicing engineering without license or experience;
• Civil rights litigation based on the violation of the defendant's rights
having been convicted by laymen using questionable engineering
evidence; and
• The use of the Racketeer Influenced Corrupt Organization (RICO)
Statutes to uncover improper training by institutes that purport to train
laymen for giving engineering testimony
A CD-ROM with abstracts of the presentations, figures, short film
clips, and animations will be distributed to the attendees at the meeting.
Accident Reconstruction, Engineering Evidence, Daubert Challenge
Forensic Histopathology, Heart, Brain
* Presenting Author
15
W12
The Medicolegal Investigation of
Recreational Diving Fatalities
James L. Caruso, MD*, U.S. Navy Undersea Medical Offices and Flight
Surgeon, Office of the Armed Forces Medical Examiner, 3929 Sweet
Briar Lane, Urbana, MD 21704; Michael D. Bell, MD*, Broward
County Medical Examiner's Office, 5301 SW 31 Avenue, Ft. Lauderdale,
FL 33312
After attending this presentation, attendees will have a basic understanding of the special physiology and specialized equipment associated
with SCUBA diving; appreciate the epidemiology of deaths associated with
recreational diving, including geographic distribution, common causes of
death, and contributing factors to these fatalities; be able to adequately
investigate and interpret the historical events and circumstantial evidence
associated with diving fatalities; understand the recommended approach to
the autopsy of persons who died while diving and be able to interpret the
anatomical findings in the context of the historical events; have handout
material and points of contact for future reference to competently investigate a diving related death; and gain a basic knowledge of other hazards
in the ocean, including dangerous marine life.
The popularity of recreational diving using scuba (self-contained
underwater breathing apparatus) has increased dramatically over the past
three decades. Present estimates place the number of active recreational
divers in the United States at between 500,000 to one million. The number
of fatalities involving U.S. citizens performing recreational dives averages
90 to100 each year. These fatalities challenge the investigators and pathologists who must investigate and certify these deaths. Recreational diving
fatalities are often litigated in civil court. This workshop is designed for the
pathologist, criminalist, attorney, and general section member who may
become involved in the investigation of a scuba diving accident or fatality.
The instruction level is intermediate.
The initial portion of the workshop will include a brief overview of
diving physiology, including the effects on the body of breathing compressed air at depth. The pathophysiology of barotrauma, nitrogen narcosis,
oxygen toxicity, gas embolism, and decompression illness (caisson disease)
will be reviewed. The pathophysiology of drowning will be reviewed since
it is a frequent final outcome in a fatal diving mishap.
Following the discussion on physiology, the epidemiology and risk
factors associated with recreational diving fatalities will be presented. A
detailed presentation on the recommended investigation of a fatal diving
mishap will be provided using illustrative cases from south Florida and the
Divers Alert Network (DAN). The importance of interviewing witnesses
and gathering information on the diver's past medical history, diving experience, pre-dive status, and the circumstances surrounding the dive will be
emphasized. The relevance of knowing the exact depth and bottom time of
the dive, as well as when and where the diver began to run into difficulty,
will be discussed. Additionally, the workshop will include a hands-on
section where typical diving equipment will be available for examination
and familiarization by attendees. This will be accompanied by a brief
discussion on the evaluation of dive gear.
In the final portion of the workshop, we will review the autopsy
protocol for scuba diving victims and emphasize those tests and observations that are helpful in determining the cause of death. The significance of
finding intravascular bubbles will be discussed, as will the proper interpretation of the findings of the autopsy. Natural diseases likely to cause sudden
incapacitation and death while scuba diving will be reviewed. Finally,
related topics such as hazardous marine animals, zoophagia, and trauma
leading to recreational diving fatalities will be presented.
Thorough handouts will be provided by the speakers, including checklists of important information to obtain regarding a diving mishap, a diving
fatality reporting form, a suggested autopsy protocol for use when diving
related fatality, and recommended resources for consultation and referral.
Cause of Death, Diving Fatalities, Medicolegal Investigation
16
W13
Facial Imaging From Human Remains
Using “Re/Face” (Reality Enhancement/
Facial Approximation by Computational
Estimation)
Kevin W.P. Miller, PhD and JoAnn Buscaglia, PhD, Federal Bureau of
Investigation, FBI Academy, Building 12, CTFSRU, Quantico, VA 22135;
Rebecca E.B. Brown* and Timothy Kelliher*, GE Global Research, One
Research Circle, Niskayana, NY 12309; Michael Taister*, FBI
Laboratory, Investigative and Prosecutive Graphics Unit, Quantico, VA
22135; Peter Tu*, GE Global Research, One Research Circle, Niskayana,
NY 12309; Wesley Turrner*, GE Global Research, One Research Circle,
Niskayana, NY 12309
The goal of this workshop is to introduce attendees to the technology
of dense facial approximation; introduce Re/Face (Reality Enhancement /
Facial Approximation by Computational Estimation); and provide
attendees an opportunity to experience Re/Face use, and provide feedback
to developers.
Re/Face is a new tool for facial approximation that will impact the
forensic community and/or humanity by significantly advancing the state
of the art. The accuracy and density of the tissue depth measurements that
are embodied in the tool provide the forensic artist with a wealth of information not previously available. Through this system, use of facial approximation as a forensic technique can be expanded into a more commonplace
tool, adding in the timely identification of individuals for law enforcement
activities. Ultimately Re/Face can be deployed into the approximation of
wartime dead leading to identification, and closure for families, of the
soldier.
Facial approximation techniques have a long history spanning over a
century. Over most of its history the techniques have advanced incrementally. Refined techniques for collecting tissue depth information and an
expansion of the database of information typify the advances in the field.
In the past ten years, the field has received heightened attention as computer-based techniques open the possibility of dramatically increased
fidelity. These techniques enable a move from a sparse set of tissue depth
information to a more robust dense set of measurements guiding the
approximation. CT scans and other medical imaging devices provide a
means for gathering tissue depth data for the entirety of a face in-vivo.
Collecting these scans, covering broad sections of the population, provide
the basis for statistical models describing total skeletal-facial variance
found within the population. Such models are used to rapidly create facial
approximations for forensic use.
Human Identification, Forensic Art, Forensic Anthropology
W14
Forensic Image and Video Processing
Zeno J. Geradts, PhD*, Netherlands Forensic Institute, Gerechtelijk Lab,
Volmerlaan 17, Rijswijk 2288 GD, Netherlands; Richard W. Vorder
Bruegge, PhD*, FBI Laboratory, 935 Pennsylvania Avenue, Room 3457,
Washington, DC 20535-0001; Jurrien Bijhold, PhD*, Netherlands
Forensic Institute, Volmerlaan 17, Rijswijk 2288 4D; Lenny Rudin, PhD*,
Cognitech, 225 South Lake Avenue, Suite 601, Pasadena, CA 911013010; Carrie M. Whitcomb, MSFS*, National Center of Forensic Science,
University of Central Florida, PO Box 162367, Orlando, FL 32816-2367
After attending this presentation, attendees will know what the possibilities are with digital images and video streams, and which techniques can
be used in forensic science.
humanity by presenting new developments in Europe and the United States
where attention is also given on aspects of accuracy.
* Presenting Author
During this workshop information will be provided on new developments of forensic investigation of (digital) images and video streams and
the use of 3-dimensional computer modeling in forensic investigations.
Traditional sources of images as evidence concern crime scene photography, and more specifically, photographs of fingerprints, tool marks,
shoe prints and other impressions. A short overview of image processing
techniques is given. Special attention is given to the introduction of artifacts
by image processing (e.g., FFT on fingerprints)
During the last 25 years the use of CCTV-camera systems has become
widespread. Typical questions concern the quality and the selection of
images from a specific camera in a multi-camera-recording. Digital processing of video streams for presentation and storage purposes, and the
compression techniques that are applied in digital CCTV-systems, lead to
questions about the integrity and authenticity of recordings.
New sources of video streams and images are video recordings from
handy cams, digital photo camera's, internet and cellular phones. Typical
questions about these recordings concern the integrity and authenticity of
the recordings, the data compression techniques used, the synchronicity of
sound and images, compensation for camera movement, and the conversion of a video stream to a higher resolution image.
We will focus on methods for digital capture and analysis of analogue
and digital multiplex surveillance recordings, state-of-the-art image
enhancement techniques as contrast stretching and de-blurring, as well as
new methods as super resolution, stabilizing and automatic tracking.
Since more images are being processed for forensic investigation, new
methods have been developed for answering questions about the interpretation of images. Examples given: Is it possible to read a license plate
number? Is our suspect, or his car, the one depicted in the image? What is
the body length of the robber or the speed of a car? Is it possible to do a
reconstruction of an accident or a shooting incident from the information in
these images? Methods for image comparison, image reconstruction, and
photogrammetry are presented and discussed. Special attention is given to
accuracy of the results and the impact on the conclusions from these investigations.
Finally, some extra attention is given to the use of 3-dimensional computer modeling in forensic investigations, since we believe that these techniques will have an impact on traditional crime scene photography.
Computer models and animations have been recently used for analyzing video by superimposition of computer generated views of the model
on the video images, for the visualization of complex scenario's in animations and for testing scenario's against video footage and evidence in crime
scene photographs. Examples: the reconstruction of car accidents from
photographs, analysis of blood spatter patterns from photographs using a
computer model of the crime scene, the visualization of wound channels in
computer models of human bodies, the reconstruction of bullet trajectories,
the reconstruction of a burglary using the limited information in dark
images from a multi camera video recording, and the analysis of firework
explosions from video recordings, photographs and geographical data.
Special attention is given to modeling techniques, the accuracy of the
models, methods for visualizing uncertainties and possibly erroneous suggestions coming from these visualizations.
Image Processing, Video, 3D Reconstruction
W15
Ephedrine: Drug or Supplement?
Ephedrine Related Compounds and
the Debate on Their Potential for
Contribution to Injury
Peter R. Stout, PhD*, Aegis Sciences Corporation, 345 Hill Avenue,
Nashville, TN 37210; Christine M. Moore, PhD*, U.S. Drug Testing
Laboratories, 1700 South Mount Prospect Road, Des Plaines, IL
60018-1804; Bruce A. Goldberger, PhD*, Department of Pathology,
Immunology & Laboratory Medicine, University of Florida College
of Medicine, 4800 SW 35th Drive, Gainesville, FL 32608; Bill J.
Gurley, PhD*, University of Arkansas for Medical Sciences, College of
Pharmacy, 4301 West Markham, Slot 522-3, Little Rock, AR 72205;
Steven B. Karch, MD*, Texas A&M University, Professor of Psychology,
Faculty of Neuroscience, 248 Psychology Building, College Station, TX
77843; Richard L. Kingston, PhD*, PROSAR, Inc., PROSAR
International Poisson Center, 1295 Bandana Boulevard, Suite 335, St.
Paul, MN 55108; Paul J. Wellman, PhD*, Texas A&M University, 248
Psychology Building, College Station, TX 77843
After attending this presentation, attendees will have a better understanding of the arguments both for and against ephedrine's potential for
harm. Data supporting these positions will also be discussed by experts in
the area of ephedrine and related compounds who have been involved in
ephedrine related litigation and policy development. Evidence from postmortem pathology, pharmacology, and analytical issues will be discussed.
Additionally, the use of AER data from poison control centers will be
discussed.
humanity by providing objective information for the forensic community
on the increasingly contentious topic of whether ephedrine and related
compounds are dangerous and should be banned or regulated or if they are
appropriately classified as nutritional supplements. This is important information for forensic toxicology professionals who are faced with interpretation of ephedrine findings and questions in court on the contribution of
ephedrine to injury and death.
With several high profile deaths recently that have had questions of
the contribution of ephedrine to the injury, many questions and opinions
about the safety and efficacy of ephedrine have been debated. The policy
landscape for ephedrine is continually evolving as congressional hearings
into the ephedra issues are currently underway. This workshop is intended
to provide a forum for the presentation of both plaintiff and defense
opinions on the safety and efficacy of ephedrine and related compounds.
This will include the issues surrounding the interpretation of ephedrine
results.
This workshop will feature a discussion on the analytical challenges
of ephedrine and related compounds in biological samples. This discussion
will also include the challenges that this presents to the interpretation of
these results. Discussion will also include mechanistic data of the understanding of the action of ephedrine on food intake and body weight. This
discussion will also include how injury may occur from a mechanistic
perspective.
As data from postmortem reports and AER data from poison control
centers have been used in the debate, discussions about the appropriateness
and use of these data will also be a part of this workshop.
An open forum period for greater discussion on opinions on ephedrine
and related compounds will also be a part of the workshop. Thorough
handouts will be provided by the speakers on data and topics presented.
Ephedrine, Injury Potential, Interpretation
* Presenting Author
17
W16
Adobe® Photoshop® for
Forensic Document Examiners
Derek L. Hammond, BA*, Forensic Document Examiner, U.S. Army
Criminal Investigation Laboratory, 4553 North 2nd Street, Forest Park,
GA 30297-5122; William J. Flynn, BS*, 7260 East Eagle Crest Drive,
Unit 33, Mesa, AZ 85207-7145; Lee Miller*
After attending this presentation, attendees will be able to move documents from files to Adobe® Photoshop®, make case file worksheets, construct court exhibits for testimony, and provide legal precedents to attorneys
and judges permitting Adobe® Photoshop® to be admissible in court.
humanity by apprizing and updating attendees of latest techniques available
to them for case work and court testimony presentation.
Courts at all levels are tightening the requirements for the admission
of expert testimony. Adobe® Photoshop® is the current state-of-the art
method for preparation of court exhibits/charts, particularly in questioned
document cases. Since the evidence processed by Adobe® is digitized, the
analyst must understand and be able to explain to what extent the images
of the evidence have been visually changed for presentation purposes.
Adobe® Photoshop® software has been available for some time but each
version adds new features. This workshop will explain the particular uses
of Photoshop® for the forensic document examiner in the areas of case file
worksheets, court exhibit preparation and the court admissibility of the use
of images processed through this software.
Digitize, Image, Photoshop®
W17
From Scene to Trial: A Quadruple
Homicide Case Study
Mary H. Dudley, MD, RN, MSN*, Sedgwick County Regional Forensic
Science Center, 1109 North Minneapolis, Wichita, KS 67214; Bradley D.
Grinage, MD*, Kansas University School of Medicine - Wichita, 1002
KU School of Medicine-Wichita, 1001 North Kansas, Wichita, KS 67214;
Diana Schunn, RN, BSN*, SANE/SART via Christi Medical Center, 3600
East Harry, Wichita, KS 67218; Kim T. Parker*, 18th Judicial Court, 535
North Main, 2nd Floor, Wichita, KS 67203
After attending this presentation, attendees will be able to describe the
national standards in crime scene investigation; differentiate between blunt
and sharp force injuries; identify the range of fire and entrance vs. exit
wounds with GSW injuries; list three key points of expert witness testimony; identify symptoms of stress & burn out; and describe the role of the
SANE nurse in fatal and non-fatal sexual assault examinations.
The national standards for death scene investigation will be presented
and related to the actual application in the multiple homicide case study
including the scene investigation and documentation, body examination
and recording. Basic principles of differentiation of blunt and sharp force
injuries with examples of each will be presented. Examples of blunt force
injuries and patterned injuries from the case study will be compared to possible weapons used to inflict injury. Medical-legal aspects of gunshot
wound injuries will include the range of fire and determining the entrance
vs. exit wounds. Photographs of the gunshot wound injuries on the six
homicide victims will be shown and detailed as presented in actual
courtroom testimony.
The forensic pathologist involved in the multiple homicide case must
attend the death scene to determine the circumstances surrounding the
death including the position and relationship of the decedents to one
another. At autopsy, the role of the forensic pathologist includes: documentation and collection of trace evidence while maintaining proper chain
of custody, external and internal examinations of the body, interpretation of
18
radiographs, collection of toxicology specimens and projectiles; and
issuing an opinion in a final autopsy report as to the cause and manner of
death.
This lecture will look at the overall care and responsibilities performed by a Sexual Assault Nurse Examiner (SANE). Health care for the
sexual assault patient has dramatically changed in the past decade. With the
development of SANE programs the specific care delivered to sexual
assault patients has focused on not only what is best for the patient but also
enhanced and equipped the healthcare provider with the appropriate education and support. The team focus has significantly improved this evolution to an advanced delivery of care. The primary goal of the sexual
assault examination is to provide a comprehensive assessment with
accurate and appropriate collection of evidence while avoiding re-traumatizing of the patient. The role of the SANE will be discussed in looking at
the patient's history, conducting a head-to-toe assessment, completing a
detailed genital evaluation and collection of forensic evidence. Courtroom
testimony will also be discussed and how the medical/legal evaluation
assists the jury in determining facts surrounding the charged case.
In a court trial the prosecutor and/or defense attorney may choose to
utilize an "expert witness." The primary role of an expert witness is to
provide the judge/jury or both, with accurate information and objective,
unbiased, and impartial opinions about issues related to the case based on
their field of expertise. The first and foremost responsibility of an expert
witness is to the court, not to the attorney or person(s) who requests their
testimony. The expert witness is used to provide their opinions impartially
despite having been hired by one of the two opposing sides in the case. The
expert witness will also be an unprejudiced educator of the judge/jury not
finding favor with the side that hired them nor with the ultimate outcome
of the case.
Critical Incident Stress Management (CISM) began as a crisis intervention model used to identify and relieve early symptoms of acute stress
in professional responders to mass casualties. Originally developed by
Mitchell (1983), techniques employed by critical incident stress debriefing
(CISD), one component of CISM, have become mainstay response to
various types of trauma throughout an array of organizations including firefighters, police and the military. The basics of CISD include structured
group discussion of the trauma associated with education of stress-related
symptoms and available resources for treatment if needed. Although CISM
has come under recent criticism regarding the lack of empirical data supporting its effectiveness, current research findings yield mixed results
regarding its ability to prevent future posttraumatic stress disorder
pathology. The workshop will present the vital roles and duties of various
members of the forensic science team stressing the importance of
cooperation and interaction of team members in a complex multiple
homicide case.
Multiple Homicides, Sexual Assault, Expert Testimony
W18
Bubble, Bubble, Boom! Fires and
Explosions in Clandestine Drug
Laboratories
John D. De Haan, MD*, Fire-Ex Forensic, Inc., 35005 Sonoma
Boulevard, Vallejo, CA 94590; Sanford A. Angelos, MS, MEd*, U.S. Drug
Enforcement Administration, 536 South Clark Street, Room 800, Chicago,
IL 60605
This workshop will provide the participants with the fundamental
information to evaluate if a fire or explosion was a result of a clandestine
laboratory. Both the arson investigator and the drug chemist will get an
insight as to the elements of each others area of expertise.
humanity by providing an excellent example of a multi-discipline approach
* Presenting Author
to a problem. Given the hazardous potential of clandestine laboratories the
workshop will alert the forensic chemist to the fire and explosion threat and
alert the arson investigators to the chemical hazards. Through interdisciplinary training the forensic examination of a fire site can lead to the
success of the investigation.
During the past several years there has been a significant increase in
the number of clandestine drug laboratories. The drug most commonly
manufactured is methamphetamine. With this dramatic increase of clandestine laboratories there has been a corresponding increase in the number
fires and explosions associated with laboratories. The very nature of a clandestine laboratory presents a variety of hazards and an increase in the possibility of a fire. The forensic chemist is increasing being called upon to
evaluate a possible clandestine laboratory site that was discovered as a
result of a fire or explosion. The arson investigator is equally being asked
to evaluate a suspected arson fire that was a clandestine laboratory site.
Providing the arson investigator with the information as to the typical
synthesis methods and the chemicals involved will great enhance their
capabilities in evaluating a fire site. The forensic drug chemist developing
an understanding of the principles of chemical fueled fires, flash fires
or solvent caused explosions will benefit greatly in processing a
laboratory site.
Historically, the typical manufacturing process for methamphetamine
was a variety of methods using phenylacetone, also known as phenyl-2propanone (P2P). These methods used P2P that was either obtained directly
or produced by the phenylacetic acid (PAA), sodium acetate, and acetic
anhydride synthesis. However, when P2P was regulated as a schedule II
controlled substance [21 C.F.R. 1308.12(g)(1)(i); 44 F.R. 71822(2/11/80)]
alternative syntheses became common. In the early 1990s clandestine
methamphetamine laboratories began to use ephedrine as the precursor.
The precursor l-ephedrine or d-pseudoephedrine is obtained commercially
from local 'super stores' and reduced to d-methamphetamine via one of the
three synthesis procedures. The first synthesis uses the chemicals
hydroiodic acid and red phosphorus; the second uses iodine and red phosphorus. The primary chemicals found at the clandestine laboratory site
using the third synthesis method are anhydrous ammonia and an alkali
metal such as either sodium or lithium. Common, to the three clandestine
synthesis process are the general chemicals of solvents, inorganic acids,
salt, and lye.
Fire science is the knowledge of fire, including the physics and
behavior of the fire or explosions. All fires are chemical in nature; however
their behavior is dependent on several factors, with fuel being a major
concern. Clandestine laboratories typically have several gallons of solvents.
The range of flammable solvents can be from diethyl ether to kerosene.
Not only do these solvents provide a source of fuel for a fire, but they also
diffuse into the air were they may ignite or explode. Most fires in clandestine laboratories are 'flash' fires caused by the flammable solvents and
their improper handling. However, the use of the water reactive alkali
metals, sodium and lithium, has introduced a new problem. A source of fuel
that can in affect self ignite. The synthesis methods that require red phosphorus introduce a fuel that once ignited will continue to burn as a chemical
reaction. This reaction has an increase in the intensity of the heat evolving
from it. This makes it an ignition source to other combustible items near it.
An additional concern is that a number of clandestine laboratory fires are
not accidental, but intentional acts of setting a fire to prevent detection.
The workshop will provide the participants with the fundamental
information to evaluate if a fire or explosion was a result of a clandestine
laboratory. Given the hazardous potential of clandestine laboratories the
workshop will alert the forensic chemist to the fire and explosion threat and
alert the arson investigators to the chemical hazards. Both the arson investigator and the drug chemist will get an insight as to the elements of each
others area of expertise.
Fire, Explosions, Clandestine Laboratories
* Presenting Author
W19
Psychological Evaluation and
Litigation of Sexual Assault Cases
Mohan Nair, MD*, 5212 Katella Avenue, Suite 106, Los Alamitos, CA
90720; Mace Beckson, MD*, PO Box 84507, Los Angeles, CA 90073;
David Gypes, JD, PhD*, 1990 South Bundy Drive, Suite 320, Los
Angeles, CA 90025; Ashok Jain, MD*, #1011,1200 North State Street,
USC/LAC, Medical Center, Los Angeles ,CA 90033; Andrew Lloyd, JD*,
1111 Town &Country Road, Suite 49, Orange, CA 92868; John
Love, JD*, Los Angeles Public Defender's Office, 9425 Penfield Avenue,
Suite 2700, Chatsworth ,CA 91311; Wesley Maram, PhD*, Sex Offender
Solutions, 1234 West Chapman Avenue ,Suite 203, Orange ,CA 92868;
Amy Phenix, PhD*, California Department of Mental Health, Sex
Offender Evaluation Program, P.O. Box 325, Cambria ,CA 93428
After attending this presentation, mental health professionals and
court officials will know the step-by-step strategies in conducting various
forms of sexual assault evaluations on both victims and defendants; will
become aware of the growing research and controversies in areas such as
clergy sexual misconduct, sexually-violent predator, profiling, eyewitness
testimony, memory distortion, suggestibility false confessions, false victimization and toxicology of date-rape drugs; will learn appropriate ways
of evaluating child sexual abuse allegations in a variety of circumstances,
i.e., child custody foster homes, schools; understand litigation strategies
from plaintiff and defense view points; and will become familiar with
instruments used for sexually violent predator evaluations, weaknesses and
controversies.
This program is intended for mental health professionals and attorneys
that deal with various areas of sexual assault. This work-up is divided up
into four sections, lasting one and one-half hours each, with questions.
GHB and related compounds are commonly used drugs within the
subculture of the "club scene.” GHB as a "date rape drug" has attracted
increasing attention by the media on the basis of high-profile cases such as
that of Max Factor heir Andrew Luster, issues of victim credibility, crime
reporting, concerns of delays in collecting urine samples, circumstances of
alleged victimization, memory problems of victims, specimen collection,
initial presentation to the emergency room, problems of forensic toxicological and laboratory methods, memory distortions, personality disorders
and of false victimization will be discussed.
Dr. Jain will review the toxicology and pharmacodynamics of gamma
amino butyric acid (GHB), Rohypnol, Chlorolhydrate and other so-called
"date-rape" drugs. Large group exposures from raves and clubs and acute
emergency room presentations of GHB will be discussed. The laboratory
analysis of GHB will be addressed. Defense and prosecution strategies in
drug facilitated rape cases will be explored.
Sexually Violent Predator Risk Assessments have been criticized as
being both unethical and unscientific. Some of these criticisms are similar
to those leveled against violent risk assessments ten years ago. The question
is, if not us, who? Dr. Amy Phenix has been involved with the California
Department of Mental Health Sex Offender Evaluation Program for the
past ten years. She will review the California experience and provide a stepby-step strategy on evaluating static and dynamic risk factors and other
tools used in the process. John Love will present the basis for the criticism
of the science and legality of the civil commitment process.
The United States Supreme Court Decision on Stogner vs. California,
notwithstanding, cases of clergy sexual misconduct, remains under intense
ongoing public scrutiny. By 1992, the Church had spent upwards of one
billion dollars for damages in medical treatment resulting from clergy
sexual abuse In addition to directly suing alleged perpetrators and the
church, lawsuits may focus in the future on institutions that have evaluated
and treated clergy offenders in an attempt to widen the defendant pool.
Even as the Stogner decision prevents criminal prosecution because of
Statute of Limitation issues, the frustration may be channeled into increased
number of civil cases, which require a lower burden of proof.
Attorneys and mental health professionals who deal with sexual
19
molestation by Clergy will learn about the steps in the forensic evaluation
of offenders and their victims. This will include utility of psychological
testing, sexual history questionnaires, instruments such as the ABEL
screening, plethysmography, polygraphic examination, and review of
treatment records. Victim evaluations will include issue suggestibility, false
memory, contamination, fabrication, and indoctrination by therapists and
others in support groups.
Assessment and litigation of child sexual assault allegations.
Presenters will review the current psychological and legal concerns in this
area. The process of developmentally appropriate examinations, appropriate and inappropriate interviewing techniques, reviewing records, psychological testing, interviewing parents and other resources. Factors
including inadequate training, evaluator bias use of anatomic dolls
contagion, parents/therapist indoctrination, and confusion, delusions,
deceptions and fabrication by the child will be covered. Comprehensive
evaluation of alleged offenders will be presented.
Sexual Assault, Litigation, Psychological Evaluation
W20
Grisly Business: An Examination of
the Modus Operandi and Signature
Characteristics of Killers and Their
Crime Scenes
Robert D. Keppel, PhD*, College of Criminal Justice, Sam Houston State
University, Box 2296, Huntsville, TX 77341-2296; Vernon Geberth, MS*,
PHI Investigative Consultants, Inc., PO Box 197, Garnerville, NY 10923
After attending this presentation, attendees will gain an understanding
of the development of Modus Operandi (MO) and Signature throughout the
history of criminal investigation; will be able to determine how the
offender's signature differs from his modus operandi from case to case
within the same series; will enable the attendees to recognize a killer's
crime scene behaviors that change and those that remain constant from one
murder to the next; and will be able to develop and construct signature and
MO testimony for court purposes.
The Grisly Business workshop will allow participants to learn that
there are crime scene indicators that relate murders even when the MO
changes. Many sexually sadistic repetitive killers, for example, go beyond
the actions necessary to commit a murder. The MO of the killer can and
does change over time as the killer discovers that some things he does are
more effective.
Beyond the MO, there are many, many killers who are not satisfied
with just committing the murder; they have a compulsion to express themselves (or do something that reflects their unique personality). The killer's
personal expression is his signature, an imprint he leaves at the scene, an
imprint he feels psychologically compelled to leave in order to satisfy
himself sexually. The core of a killer's signature will never change. Unlike
the characteristics of an offender's MO, the core remains constant.
However, a signature may evolve over time in some cases where a
necrophilic killer performs more and more post mortem mutilation from
one murder to the next.
Experts once described the nature of the signature as the person's
violent fantasies, which progress in nature and contribute to thoughts of
committing extremely violent behavior. As a person fantasizes over time he
20
develops a need to express those violent fantasies. Most serial killers have
been living with their fantasies for years before they finally bubble to the
surface and become translated into behavior. When the killer finally acts
out, some characteristic of the murder will reflect some unique aspect
played over and over in his fantasies.
Likewise, retired NYPD homicide detective Vernon Geberth (1996)
wrote that it's not enough just to consummate the murder, the killer must act
out his fantasies in some manner over and beyond inflicting death-producing injuries. This is the signature of the killer.
Detectives who investigate a series of murder scenes look for the same
type of extraordinary violence and a bizarre set of similarities. Their gut
instincts tell them there's more here that's alike than different, but another
homicide investigator will say the killer used a pipe wrench as a blunt
instrument here, a hammer there, and in this third crime we can't even
figure out what weapon was used. Maybe he draped a pair of underpants on
the victim's left leg. In the next crime scene, the underpants were on her
right leg or maybe still on the bed. Yet, in each case, the victim was obviously beaten well beyond the point of death by an assailant whose violence
seemed to increase in frenzy while he was attacking her. Also, the killer
seemed preoccupied with the victim's clothing and took some time to
arrange the crime scene even though there might have been people living
just upstairs. These are the psychological calling cards the killer actually
needs to leave at each scene. Other examples of signatures covered in the
workshop are mutilation, overkill, carving on the body, leaving messages,
rearranging or positioning the body, engaging in post-mortem activity, or
making the victim respond verbally in a specified manner. These constitute
a signature behaviors of a particular killer. What is important about a killer's
signature, then, is that killers learn to treat victims the way they do in their
fantasies, always attempting to satisfy their fantasies as they move from one
victim to the next.
Workshop participants will learn that identifying a killer's signature is
a multidisciplinary effort, using the findings of medical examiners, anthropologists, dentists, criminalists, and behavioral scientists. An examination
of a series of murders requires typically, the information from the police
investigative file, which includes officer's reports, statements, crime laboratory reports, crime scene diagrams, photographs, videotapes of crime
scenes, autopsy reports, and report of other experts.
Police investigators and prosecutors need cases linked for their own
purposes. From an investigative standpoint, the linking of crimes enables
investigators to pursue the same suspect instead of operating without the
knowledge that the cases were linked. Prosecutors want similar cases
linked so the defendant can be tried on multiple charges in the same trial.
The threshold for using MO and signature as evidence at trial differs
from state to state. In Virginia, for example, the Supreme Court in the
Timothy Spencer murder convictions held that: "evidence of other crimes,
to qualify for admission as proof of modus operandi, need not bear such an
exact resemblance to the crime on trial as to constitute a 'signature,' but it
is sufficient if the other crimes bear a singular strong resemblance to the
pattern of the offense charged and the incidents are 'sufficiently idiosyncratic to permit an inference of pattern for purposes of proof,' thus tending
to establish the probability of a common perpetrator."
The Grisly Business workshop presentations will highlight those
cases most of which have had courtroom testimony and/or reports written
dealing with MO and Signature aspects: Those cases are Morris Frampton,
George Russell, and Robert Yates (Washington State), Richard Cottingham
(New York), Timothy Spencer (Virginia), Terrence Burlingham (British
Columbia, Canada), and David Parker Ray, the New Mexico Torture
Chamber Killer.
Modus Operandi, Signature, Murder Characteristics
* Presenting Author
W21 Y-STR Analysis on Forensic Casework
Sudhir K. Sinha, PhD*, Reliagene Technologies, 5525 Mounes Street,
Suite 101, New Orleans, LA 70123; John Ballantyne, PhD*, Department
of Chemistry, University of Central Florida, Building #5, 400000 Central
Boulevard, Orlando, FL 32816-2366; Charles E. Barna, BS*, Michigan
State Police, 714 South Harrison Road, East Lansing, MI 48823; John M.
Butler, PhD*, National Institute of Standards and Technology, 100 Bureau
Drive, Mail Stop 8311, Gaithersburg, MD 20899-8311; Ranajit
Chakraborty, PhD*, Center for Genome Information, Department of
Environmental Health, University of Cincinnati, College of Medicine,
3223 Eden Avenue, Cincinnati, OH 45267; Meihua Chu, PhD*, MiraBio,
Inc., 1201 Harbor Bay Parkway, Suite 150, Alameda, CA 94502; Peter de
Kniijff, PhD*, Forensic Laboratory for DNA Research, MCG-Department
of Human and Clinical Genetics, Leiden Medical Center, PO Box 9503,
Leiden, 2300 RA, The Netherlands; Paul A. Foxall, PhD*, Applied
Biosystems, 850 Lincoln Center Drive, Foster City, CA 94402; William E.
Frank, MS*, Illinois State Police, R & D Laboratory, 2060 Hill Meadow
Drive, Springfield, IL 62702; Leonor Gusmao, PhD*, Institute of
Pathology and Immunology for the University of Porto, R. Dr. Roberto
Frias, s/n, Porto 4200-465, Portugal; Kimberly A. Huston, BS*, Genetic
Identity, Promega Corporation, 2800 Woods Hollow Road, Madison,
WI 53711; Carll Ladd, PhD*, Connecticut Forensic Laboratory,
Forensic Biology Unit, 278 Colony Street, Meriden, CT 06451; Barbara
Llewellyn, PhD*, Illinois State Police, R & D Laboratory, 2060 Hill
Meadow Drive, Springfield, IL 62702; Timothy P. McMahon, PhD*,
Armed Forces DNA Identification Laboratory, Armed Forces Institute of
Pathology, 1413 Research Boulevard, Building 101, 2nd Floor,
Rockville, MD 20850; Alan J. Redd, PhD, University of Arizona, Division
of Biotechnology, Biological Sciences West 239, Tucson, AZ 85719;
Reena Roy, PhD, St. Louis Police Crime Laboratory, 111 South
Merammec, Clayton, MO 63105; Jaiprakash G. Shewale, PhD*,
ReliaGene Technologies, Inc., 5525 Mounes Street, Suite 101, New
Orleans, LA 70123
Upon completion of this workshop, the participants should be able to
understand the characteristics of the polymorphic markers on the
Y-chromosome, and population genetics. The participants should also
become familiar with multiplexed Y-STR systems and their usefulness and
limitations in forensic casework.
Upon attending this workshop, the forensic community will obtain the
knowledge on the usefulness of Y-STR analysis in forensic casework. The
participants will learn the aspects of the structures of different Y-STR
markers, Y-STR markers used in forensic applications, population genetics,
laboratory validation requirements, result interpretation issues and new
Y-STR markers on the horizon.
Y-Chromosome Short Tandem Repeats (Y-STRs) have become a very
useful tool in forensic casework, paternity testing and male lineage studies.
Different types of challenges can arise in the analysis of autosomal STRs,
mainly in the interpretation of results for samples containing mixtures of
male and female DNA. The Y-STRs can be utilized in resolving such cases.
The use of Y-STRs makes it possible to obtain an exclusive profile of male
DNA in a sample containing mixtures of male and female DNA.
The contents of the workshop will target forensic scientists who are
familiar with STR analysis and are interested in implementing or who have
implemented Y-STR analysis for forensic casework. The Scientific
Working Group on DNA Analysis Methods (SWGDAM) has identified a
core set of eleven Y-STR loci for forensic analysis in the United States.
Several new Y-STR loci have been discovered in the past few years.
The presentation topics will cover aspects of Y-STR analysis in
forensic casework including the characteristics of Y-STRs, multiplex
systems, validation studies according to the DAB standards, database
studies, result interpretation, population genetics, courtroom testimony,
* Presenting Author
QA/QC issues and casework studies. The speakers will include international scientists who have made milestone contributions, and scientists
from academic institutions, crime laboratories, government institutions and
private industries active in research and development of Y-STRs. Thus, a
unique combination of expertise will be available for interpretation and
discussion.
Y-STRs, Y-Chromosome, Forensic Casework
W22
Recovery, Examination, and Evidence of
Decomposed and Skeletonized Bodies:
An Anthropological and Entomological
Approach
M. Lee Goff, PhD*, Professor and Chair, Forensic Science Program
Chaminade University, 3140 Waialae Avenue, Honolulu, HI 96816-1578;
Wayne D. Lord, PhD*, Federal Bureau of Investigation, NCAVC, FBI
Academy, Quantico, VA 22135; Edward T. McDonough, MD*, Office of
the Chief Medical Examiner, 11 Shuttle Road, Farmington, CT 06032;
William C. Rodriguez III, PhD*, Office of the Armed Forces Medical
Examiner, AFIP, 1413 Research Boulevard, Building 102, Rockville,
MD 20850
After attending this presentation, attendees will be able to recognize
bioenvironmental evidence; properly collect and preserve such evidence;
and record supplementary data needed for later analyses of bioenvironmental evidence.
humanity by presenting techniques from the disciplines of anthropology,
entomology and pathology that can be employed in the processing of crime
scenes. This interdisciplinary approach will maximize the information
obtained from bioenvironmental evidence gathered from a scene.
Techniques will be presented which, if used, will assure the proper
collection, preservation and documentation of these types of evidence for
later analyses.
One of the most challenging cases faced by any forensic scientist or
investigator is that of the badly decomposed or skeletonized body. It is a
common misconception that such remains, particularly those in a field or
wooded area, provide little useful information concerning the circumstances of death. However, through the applications of techniques from the
fields of anthropology and entomology, significant data may be obtained.
The outdoor death scene is quite unique, since the remains and associated
evidence can be viewed as temporary alterations to the ecology of the
immediate area. Methods and techniques for the recognition, collection,
preservation, and interpretation of this "bioenvironmental" evidence will be
presented during the workshop. This workshop is designed to be at an intermediate level, with an overview of anthropological and entomological
techniques, followed by considerations of recent advances in these areas of
research combined with techniques for processing outdoor crime scenes.
Decompositional processes will be covered along with the varied applications of entomological evidence, including entomotoxicology, preservation
and processing of entomological evidence, and applications of entomological evidence in cases involving the living as well as the dead.
Entomology, Anthropology, Postmortem Interval
21
W23
Application of the Principles of
Pharmacology and Pharmacokinetics to
the Interpretation of Drug Blood Levels
David M. Benjamin, PhD*, 77 Florence Street, Suite 107, Chestnut Hill,
MA 02467-2121; Timothy P. Rohrig, PhD*, Regional Forensic Sciences
Center, 11009 North Minneapolis Street, Wichita, KS 67214; Adam
Negrusz, PhD*, Department of Pharmacodynamics; University of Illinois,
833 South Wood Street, Chicago, IL 60612; Robert H. Powers, PhD*,
Hamilton County Coroner's Office, 3159 Eden Avenue, Cincinnati, OH
45219; Dorothy E. Dean, MD*, Franklin County Coroner's Office, 520
King Avenue, Columbus, OH 43201
After attending this program, forensic scientists will be able to analyze
drug blood level data from a pharmacokinetic perspective; be able to
demonstrate an understanding of the four pharmacokinetic phases of drug
disposition: absorption, distribution, metabolism and excretion; be able to
recognize mechanisms of potentially toxic drug interactions and additive
toxicities, will avoid pitfalls in interpreting drug blood level data involving
postmortem blood samples, and will be able to recognize the variability
involved in obtaining post-mortem blood samples from cardiac and
peripheral sites.
Forensic scientists and clinicians alike are frequently called upon to
interpret the results of blood and urine drug analyses in order to determine
the cause of toxicity or death. With some drugs, there is a good correlation
between their concentrations in blood and their therapeutic, toxic and lethal
levels. With other drugs, there is no such relationship, and with still others,
it is the unbound or unconjugated drug concentration that correlates well.
Inhibition or enhancement of any of the four pharmacokinetic phases:
absorption, distribution, metabolism and excretion can increase toxicity of
a given dose or provide an increased safety margin.
Many errors are made by physicians, pathologists, and toxicologists
who lack formal training in pharmacokinetics, regarding the interpretation
of drug blood levels, especially samples taken in the post-mortem period.
Few courses exist to teach the required pharmacokinetic skills to forensic
scientists, thus perpetuating the same frequency of errors. This symposium
is designed to teach many of the skills required to properly interpret drug
blood levels obtained from living and deceased individuals. The course
begins with a review of the essential pharmacokinetic relationships
required to understand drug disposition and continues with presentations of
cases selected to demonstrate the application of the principles of pharmacology and pharmacokinetics to the interpretation of drug blood levels in
living and deceased subjects. Only few courses have been designed to teach
these essential skills and attendees are sure to gain valuable training in the
application of pharmacokinetics to their work.
Pharmacokinetics, Interpretation of Drug Blood Levels, Postmortem
Redistribution
WS1 Pediatric Death Investigation Including
Subtle Forms of Fatal Abuse: From the
Scene to the Courtroom
Karen F. Ross, MD*, Assistant Coroner, Jefferson Parish, 2018 8th Street,
Harvey, LA 70058; Joni L. McClain, MD*, Medical Examiner, Dallas
County Medical Examiners Office/SWIFS, 5230 Medical Center Drive,
Dallas, TX 75235; Kathleen A. Diebold, BA*, Child Death Specialist, St.
Charles, Jefferson, and Franklin Counties Medical Examiner, 1402 South
Grand Boulevard, St. Louis, MO 63104; Robert G. Williams, DDS*, Chief
Forensic Odontologist, Southwestern Institute of Forensic Sciences,
11661 Preston Road, Dallas, TX 75230; John W. Thompson, Jr., MD*,
Associate Professor of Clinical Psychiatry, Tulane University School of
Medicine, Dept. of Psych & Neurology TB 53, 1440 Canal Street, 10th
Floor, New Orleans, LA 70112
Upon completion of this workshop, the participant will be able to recognize the unique aspects of investigating deaths in childhood; be familiar
with autopsy procedures and investigative techniques which may be
employed in such cases; be aware of potential problems and possible solutions in pediatric toxicology; be able to recognize subtle forms of fatal child
abuse including neglect; understand that designation of manner of death as
accident or undetermined does not exclude the possibility of prosecution of
the alleged perpetrator; recognize the role of the forensic odontologist in
identification and prosecution of certain cases, and become familiar with
the psychological features of the perpetrators of filicide including possible
motivating factors.
Investigation of sudden unexpected deaths in childhood presents
unique challenges to all involved in the process from the scene investigator
to the forensic pathologist. In many cases, the cause of death is obvious as
is the manner whether natural, accident, or homicide. In other cases,
injuries identified at autopsy may obviously be the result of non-accidental
injury; however, identification of the perpetrator may be difficult. In still
other cases, the death, while classified as accidental or undetermined, may
be the result of negligence on the part of the caretaker who may still be
prosecuted. Thorough investigation, from the scene to the morgue, and
cooperation between the various investigating agencies and representatives
of the death investigation system ensure that cases of fatal child abuse are
recognized and that the perpetrator is identified and prosecuted. This same
cooperation may prevent innocent people from being wrongly accused of
and charged with such crimes. The training of people specifically to investigate child hood deaths has been initiated in some medical examiner
systems. Though there really is no sub-sub-specialty of pediatric forensic
pathology, many forensic pathologists, for varying reasons, become particularly interested in childhood deaths. The special interests of trained professionals can only be considered a positive step in childhood death investigation. In addition to those involved in the primary aspect of the investigation, other forensic specialists such as odontologists and psychiatrists
may be involved in the investigation of these cases, specifically in the identification and prosecution of the perpetrators. This workshop emphasizes
the role of each of these professionals in the successful investigation of
childhood deaths through lectures and case presentations.
Pediatric, Death, Investigation
22
* Presenting Author
WS2 Andrea Yates Trial —
Revisited by the Defense
Emanuel Tanay, MD*, Clinical Professor of Psychiatry, Wayne State
University, 2977 Philadelphia Drive, Ann Arbor, MI 48103; Lise Van
Susteren, MD*, Assistant Clinical Professor of Psychiatry, Georgetown
University, 4707 Conn Avenue, Washington, DC 20008; George
Parnham*, 440 Louisiana Street, Suite 800, Houston, TX 77002
The goal of this presentation is to inform the participants about infanticide, insanity defense, and the ethical role of forensic psychiatry in
criminal trials.
humanity by informing members on the usefulness of insanity defense;
increasing awareness of the conflict between requirements of law and
science; highlighting the distortions that occur as the result of prosecutorial concerns with public opinion; and last not least, increasing
awareness that a criminal trial is about justice and not gamesmanship.
A sane mother cannot will herself to kill her children, nor can she will
herself not to when driven by psychotic delusions. To present the killing
of five children by a mother as a result of a sane intellect is unreasonable
and yet that is what the prosecution had to argue in order to obtain firstdegree murder conviction.
The killing of five children by a mother is not a defect of conscience
but a psycho-biological derangement. Maternal instinct is not only a
matter of psychology but of biology. The mother-child bond is instinctual,
therefore, the killing of five children by a human mother is a psycho-biological aberration. Yet, the prosecutors and their expert claimed that
Andrea Yates drowned her children because "The children had become a
hindrance, and she wanted them gone." The psychiatric expert witness
who testified against insanity defense in the Andrea Yates case was paid
by taxpayers $105,636.99.
One wonders what social value justified this type of expenditure on
the part of the prosecutor's office. A psychotic mother who drowned her
* Presenting Author
five children would be confined for the rest of her life if she was found
insane or guilty of first degree murder. Thus, the effort from the persecution's perspective was about the location of the life-long confinement of
this tragic figure.
The Congress was outraged by the insanity verdict in the John
Hinckley case and crafted legislation to prevent such "abuse." In reality
insanity pleas were made in only 2% of felony cases and failed in more
than 75% of the cases. Most severely mentally ill homicide defendants are
found guilty of first degree murder under the wrongfulness test.
Andrea Yates a chronically psychotic woman drowned her five
children in a bathtub and called the police. She was found not insane and
guilty of first-degree murder. There were no legislative hearings, no
protest from organized medicine or psychiatry. John Hinckley like Andrea
Yates was chronically schizophrenic. Neither Hinckley nor Yates received
appropriate treatment before the criminal offense.
The Andrea Yates case and the many other killings of children by
psychotic mothers raise profound moral, legal and psychiatric issues.
Should we judge human behavior by its consequences and disregard the
state of mind that gave rise to this behavior? Should we have a criminal
justice system which does not consider intent? Are we to disregard the
delusional beliefs that give rise to homicidal behavior? Should we make
no distinction between rational beliefs and beliefs that are symptoms of
psychosis? Are we to assume that there was no causal link between the
psychosis and the drowning of five children?
The principle that knowledge of the difference between right and
wrong is the criterion for insanity is the law. It cannot be questioned by an
expert witness. Nevertheless, a psychiatric witness has to make the
concept meaningful within his or her science. The very fact that psychiatric expert testimony is admissible as a matter of law presumes that the
testimony will be given in accordance with the principles of psychiatry
and pass the "Frye Test." It is our contention that relying upon a delusion
as proof for knowledge of wrongfulness is contrary to science, law, and
common sense.
Insanity, First-Degree Murder, Ethics of Expert Testimony
23
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DALLAS
B1
Criminalistics
Applications of Forensic
Astronomy in Turkey
H. Bülent Üner, PhD*, Institute of Forensic Sciences, Istanbul
University, Adli Týp Enstitüsü, Cerrahpaþa, Istanbul, 34301,
Turkey; Emre Albek, MD, Cerrahpaþa Medical Faculty, Istanbul
University, Cerrahapaþa Týp Fakültesi, Istanbul, 34301, Turkey;
Ismail Cakir, PhD, Council of Forensic Medicine, Turkey, Adli Týp
Kurumu, Cerrahpaþa, Istanbul, 34246, Turkey
After attending this presentation, attendees will understand applications of forensic astronomy.
Frequently, astronomy proves to be an important contributory
science to criminalistic investigations.
Crime scene investigation on the date and at hours calculated by
forensic astronomers is the only means to ascertain whether conditions
of visibility at the time of a crime are consistent with the information
obtained from victims, eye witnesses and the alleged assailants. During
the crime scene investigations, meteorologic conditions at the time of the
alleged crime should also be taken into consideration.
The forensic astronomer using degree of longitude and latitude of
the scene as well as data from regular astronomical almanacs, calculates
the moon phase, the time of moonrise and moonset, the time of sunrise
and sunset, so determines the nights on which the same lunar illumination as in the night of the event will be present
In Turkey with its almost wholly muslimic population, persons
involved in a criminal case sometimes report the time of occurrence in
association with the muezzine’s call for prayer. In such cases, forensic
astronomers have to calculate the exact hour of the praying call at the
time of the alleged crime.
Criminalistics, Astronomy, Sun and Moon
B2
A Rapid Analysis of Low Explosives
With SEM/EDS—A Case Report
Ismail Cakir, PhD*, Council of Forensic Medicine, Turkey, Adli Týp
Kurumu, Cerrahpaþa, Istanbul, 34246, Turkey; H. Bülent Üner, PhD,
Institute of Forensic Sciences, Istanbul University, Adli Týp Enstitüsü,
Cerrahpaþa, Istanbul, 34301, Turkey; Salih Cengiz, PhD, Institute of
Forensic Sciences, Istanbul University, Adli Týp Enstitüsü,
Cerrahpaþa, Istanbul, 34301, Turkey
After attending this presentation, attendees will understand rapid
analysis of low explosives with SEM/EDS.
Scanning Electron Microscopy Energy Dispersive X-Ray Analysis
(SEM/EDS) is a very useful method to analyze for most of inorganic
materials like gunshot residues, paints, soil, glass etc. Also it is possible
a rapid analyze of low explosives with SEM/EDS.
In the present study, we carried out rapid analysis of low explosives
(potassium chlorate and sulphur) by the method mentioned above.
While four young men were at the seaside for fishing, they wanted
to prepare a device of explosive-fishing by adding a powdered material
to another that was in a bottle. At this time the bottle exploded with a
huge noise and four young men were injured.
Crime scene investigators found two different powder materials
and pieces of a broken glass bottle at the scene.
24
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2004
Although in this case, it was believed by crime scene investigators
that these materials found at the scene, were calcium carbide, they were
sent by the public prosecutor for analysis to our laboratory.
One of these materials was slightly moistened, white powder, and
the other was dry, yellow powder. Approximately two grams of each
powder was placed and heated in an 110º C oven for 1 h. Then materials
removed from oven and were allowed to cool to room temperature. Each
of these materials was placed on two different double-sided adhesive
tape then attached to a stub, and coated with carbon. SEM/EDS analyses
for these materials were carried out using a JEOL 5600LV scanning
electron microscope equipped with a LINK-ISIS 300 X-ray analyzer.
The results obtained in this study indicate that elemental composition of the white powder consisted of potassium, chlorine, and oxygen
(its compound was potassium chlorate), yellow powder was sulphur.
SEM/EDS, Low Explosives, Potassium Chlorate and Sulphur
B3
Evaluation of Y-STR Markers
Using a Cajun Population
Katherine L. Cross, BS*, National Medical Services, 3701 Welsh Road,
Willow Grove, PA 19090; Ranajit Chakraborty, PhD, University of
Cincinnati, 3223 Eden Avenue, Kettering Lab, Room 110, University of
Cincinnati Medical Center,
PO Box 670056, Cincinnati, OH 45267-0056; Mary T. Jablonski, BS,
MS, National Medical Services, 3701 Welsh Road, Willow Grove, PA
19090
Do the eight minimal haplotype Y-STR markers provide enough
discrimination in a population that is potentially highly related? In this
study, it can be demonstrated that in related populations, eight markers
do not provide enough discrimination of the individuals in that population.
This presentation will impact the forensic community by demonstrating how the eight minimal haplotype Y-STR markers alone may be
insufficient to reach individual characterization; therefore, additional
markers may need to be used to discriminate some populations.
The Cajun population of southern Louisiana has a long history. The
original population consisted of only a few family units that were
continuously displaced from France to Canada to Louisiana. This
population had sustained geographical and social isolation from the
1700s until the mid-late 1900s. Given the history of the Cajuns and
since Y-STR’s are inherited virtually unchanged from father to son(s),
the possibility of observing a minimal number of different profiles form
the Cajun population is greater than in the general population. Samples
from the southern Louisiana area Cajun and non-Cajun populations were
collected and processed. These samples were organically extracted,
quantified, amplified, and analyzed using capillary electrophoresis. The
eight minimal haplotype STR’s plus two additional Y-STR’s were analyzed. Even with ten loci, the results show more similarities and exact
matches in the Cajun population than in a non-Cajun population.
Y-STR, Related Population, Discriminating Power
* Presenting Author
B4
Optimization of Spermatozoa Capture
During the Differential Extraction
Process for STR Typing With the
Potential for Automation
Pamela L. Marshall, MS*, Maryland State Police Crime Laboratory,
1201 Reistertown Road, Pikesville, MD 21208; Art Eisenberg, PhD,
University of North Texas Health Science Center, 3500 Camp Bowie
Boulevard, Fort Worth, TX 76107
After attending this presentation, attendees will learn about a new
method of spermatozoa capture that could enhance the differential
extraction process already in place in most crime laboratories and learn
about the research involved in this method
This research project shows tremendous potential as a new and
improved method for obtaining a complete and separate sperm fraction
from sexual assault samples. It could lead to an automated method,
establishing a faster technique for the differentiation of sexual assault
samples, and could be a tremendous breakthrough in the backlog of
cases most states are currently facing.
In 1998, within the United States, it is estimated that a rape
occurred every 2.3 minutes. In 1995, according to the Bureau of Justice
Statistics, an estimated 350,000 rapes or sexual assaults (R/SA) were
experienced by persons age 12 or older. Of the estimated 100,000 R/SA
reported, there were only approximately 25,000 cases analyzed by crime
labs nationwide. The majority of crime laboratories throughout the U.S.,
especially those in major metropolitan cities, have a significant backlog
of unresolved R/SA cases. With the implementation of the Convicted
Offender Database (CODIS), it is essential that all R/SA cases be analyzed, especially those lacking a known suspect. The comparison of the
short tandem repeat (STR) profiles derived from sperm DNA recovered
from evidentiary material with CODIS samples would provide the police
with critical investigative leads resulting in the identification of the
assailant.
The goal of this research is to develop a cellular sorting method for the
isolation of the sperm cells from sexual assault samples which will: 1) take
advantage of additional differentiating cellular features (extracellular antigenic sites) so that the separation of the cell types will be complete, 2)
provide a more efficient means of sperm recovery, and
3) utilize commercially available equipment for the automation of the entire
process, resulting in a cost effective and more reliable process. Overall, the
proposed technique will increase the probability of success in the analysis
of sexual assault case samples. (NIJGrant #: 2000-IJ-CX-K009).
Differential Extraction, Sexual Assault, Short Tandem Repeats
B5
Carboxyhemoglobin Analysis
in Gunshot Wounds
Hakan Kar, MD, The Council of Forensic Medicine of Turkey, Adli Tip
Kurumu, Cerrahpasa, Istanbul, 34246, Turkey; Salih Cengiz, PhD*,
Institute of Forensic Sciences, Adli TIP Enstitüsü Cerrahpaþa,
Istanbul, Turkey; Bülent Üner, PhD, Institute of Forensic Sciences, Adli
Týp Enstitüsü Cerrahpaþa, Istanbul, 34300, Turkey; Harun Tugcu,
MD, Gülhane Military Academy of Medicine, Department of Forensic
Medicine, Gülhane askeri Týp Akademisi, Ankara, Turkey; Necmi
Cekin, MD, The Council of Forensic Medicine of Turkey, Adli Tip
Kurumu, Cerrahpasa, Istanbul, 34246, Turkey
After attending this presentation, attendees will understand the definition of the entrance and exit wounds together with estimation of the
shooting range and the major criteria in determination of the origin of the
gunshot wounds.
* Presenting Author
The goal of this Presentation is the definition of the entrance and
exit wounds together with estimation of the shooting range and the major
criteria in determination of the origin of the gunshot wounds.
Today, usage and ownership of firearms is rising rapidly over the
world. As a result of this increasing demand, the rate of usage of firearms
in crimes is also rising, even in countries where the ownership is strictly
under the control of the government.
Determination of the origin in gunshot wounds is a process that
starts with crime scene investigation and judicial investigation findings,
developing with examination or autopsy together with laboratory tests
and finalizing with jurisdiction. It’s mentioned that the definition of the
entrance and exit wounds together with estimation of the shooting range
are the major criteria in order to determine the origin of the gunshot
wounds. Crime scene investigation, collecting the evidences those are
related to the case (like the guns, bullets, fingerprints, gunshot residues,
clothes with gunshot holes, etc.), physical and morphological properties
of the gunshot wounds, detection of the gun powder and soot on and
under the dermis during the autopsy, analysis of the gunshot residues on
the clothes and hands with chemical examinations, applying some
advanced technological tests like “Neutron Activation Analysis,”
“Atomic Absorption Spectrophotometers,” “Scanning Electron and Ion
Microscopy,” “Imaging Analysis” are enabling us to determine the
entry-exit wounds and firing range.
Getting the clothes after washing and cleaning for the tests, clothes
can mould and become useless because of the improper transportation
and storage conditions. Even in some cases clothes that should be kept
as evidences are being lost at crime scenes or hospitals during first aid.
In some cases gunshot wounds become impossible to examine
when the body has been washed or embalmed, operated medically or
cleaneased to hide the remains. There may be indeterminate results in
cases when the epidermis is decayed, scar tissuehas formed, or insect
activities occur around the gunshot wounds.
Determination of shooting range and entry-exit wounds cannot
always be possible with classical and advanced methods mentioned
above. In such cases carboxyhemoglobin (COHb) analysis in the blood
samples obtained from the muscle tissues right under the gunshot wound
will be a helpful criteria to determine entry-exit wounds.
The aim of this study is to compare the carboxyhemoglobin levels
in blood samples obtained from the muscle tissues and veins in contact,
near contact and close range gunshot wound cases; and with this method
to distinguish the gunshot wounds from other types of wounds, to differentiate the entry and exit wounds and to define if the firing range is
contact or not.
Material and Method: 52 contact, near contact and close range
gunshot wound autopsy cases examined at the State Institute of Forensic
Medicine between May 2001 and May 2002 were included. COHb
levels of blood expressed from the muscle tissues at the entry and exit
sites and veins were analyzed. The blood samples were filtered with the
“Cloth Catcher” and analyzed with “Co-Oximeter.”
Findings: 39 of the cases were shot with handguns, 13 with
shotguns; and 47 of the cases were shot from contact range, 2 from near
contact range and 3 from close range. The difference between the
average COHb levels of the entry wounds (handgun: 8,673, shotgun:
23,000) and the exit wounds (handgun: 4,655, shotgun: 15,871) were
statiscially significant in both handgun and shotgun wounds. And differences between the average COHb levels of the entry, exit wounds and
venous blood samples (handgun: 1,545, shotgun: 5,729) were also statiscially significant.
Discussion and Conclusion: As a result; in suspected gunshot
wound cases, a significant difference between the COHb levels of the
venous blood and the blood expressed from the muscle tissues at the
wound sites suggests that the lesion is a gunshot wound and is a result of
a contact range shot. Therefore this method will be helpful criteria in differantial diagnosis of the suspected or controversial gunshot wound
cases when used with other diagnostic criteria.
Carboxyhemoglobin, Gun Shot, Wounds
25
B6
Advancements Pertaining to
the Physical Developer Process
B8
Heteroplasmy Pattern in D-Loop
Region of Mitochondrial DNA
David E. Burow, BS*, Marshall University, 1401 Forensic Science Drive,
Huntington, WV 25701; Don Seifert and Antonio A. Cantu, PhD, United
States Secret Service, 950 H Street NW, Washington, DC 20001
Chong Min Choung, Zhe Li, Yoon Seong Lee, Jung Bin Lee, and Soong
Deok Lee*, Department of Forensic Medicine, SNU Medical School,
28 Yongon-dong Chongno-gu, Seoul, 110-799, Korea
After attending this presentation, attendees will understand how
latent fingerprints are developed on porous surfaces with the use of
physical developer.
This presentation will demonstrate the reduction in cost of physical
developer along with improved latent fingerprint development.
The silver physical developer (Ag-PD) is a water-based reagent that
is commonly used to develop latent fingerprints on porous surfaces. It
usually follows DFO and/or ninhydrin in the processing of latent prints.
The Ag-PD reacts with certain water-insoluble components of latent
print residue (e.g., lipids) while both DFO and ninhydrin are solventbased reagents that react with certain water-soluble components (e.g.,
amino acids). The Ag-PD works by depositing silver on “triggering”
(catalytic) sites in the latent print residue. The Ag-PD consists of silver
nitrate, citric acid, surfactants, and a mixture of ferrous and ferric salts,
of which the ferrous ions act as reducing agents for the silver ions.
Recent changes were made to the Ag-PD when highly purified water was
substituted for distilled water. These changes resulted in better performance and a reduction in components used and thus, a reduction in cost.
This presentation will cover the approach taken in making the changes
and the improvements they made.
After attending this presentation, attendees will learn about the heteroplasmy pattern in D-loop region of mitochondrial DNA.
This presentation will impact the forensic community by demonstrating how heteroplasmy pattern in the D-loop would help when interpreting the sequencing results for the mitochondrial DNA.
Mitochondrial DNA (mtDNA) is polymorphic in the D-loop region.
This polymorphism is used in various areas, among which individual
identification in forensics and confirmation of maternal lineage are the
common usuage. The mtDNA is small and circular, and exists as a lot of
copy number even within a cell. With these characteristics mtDNA has
become a valuable tool in individual identification, especially when the
conventional typing for the autosomal STRs are unavailable such as in
old degraded samples, bone and nail.
Several may be obstacles when using mtDNA. Heteroplasmy is one
of those. There may exist different types of mtDNA within a cell or even
within a mitochondrion. When someone performs mtDNA sequencing,
heteroplasmy must be considered. It may be difficult to tell case of different origin from case of same origin showing heteroplasmy.
Conversely the insight into the heteroplasmy may increase the discrimination power of mtDNA typing or may give some idea for how the polymorphism of mtDNA occurs.
Several have reported about cases of heteroplasmy, but these do not
seem to be enough. We have screened 433 maternally unrelated Koreans
using DGGE and have found 82 cases of heteroplasmy in HVI. We have
sequenced all the samples and decided how heteroplasmy exists. The
pattern of heteroplasmy and several related results including heteroplasmy pattern among different organs within an individual or age relationship will be presented.
Fingerprints, Physical Developer, Porous Surfaces
B7
Statistical Process Control in the
Interpretation of DNA STR Profiles
Lawrence A. Presley, MS, MA*, National Medical Services, 3701 Welsh
Road, Willow Grove, PA 19090
The goal of this presentation is to describe a data-driven method for
the interpretation of low level DNA samples.
In 1992, the National Research Council recommended the use of the
“ceiling principle” as a “conservative” estimate of the likelihood of a random
match. This assumption was based on the limited availability of data, and by
1996 the National Research Council suggested that this “conservative”
estimate was no longer necessary. Currently, many forensic DNA interpretational guidelines suggest that rfu cutoffs of 100 or 150 rfus, are a “conservative’ cutoff estimate of what are callable DNA peaks. This “conservative”
estimate is generally based on pre-casework validation studies of low-level
DNA samples, but a continuous process may offer a better estimate.
Statistical process control offers a better and more data-driven
approach for the estimation of the range of callable DNA peaks in a low
level sample, rather than a pre-determined single number estimate. It is
not scientifically reasonable to suggest a peak of 151 rfus is conclusively
callable while a peak of 149 rfus is not without some continual basis for
comparison. Statistical process control provides a reliable and accepted
means of establishing an estimated range of rfu values for low-level DNA
peaks, and a historical, data-driven, and continuous approach for interpreting low-level DNA peaks.
Background negative control sample data were collected, and meaningful noise levels were established. The variation ranges around the noise
levels offer a degree of confidence around a recorded value, and suggest a
range of values that can be used for the interpretation of ‘real’ low-level
DNA peaks. Both positive and negative control samples were run with
several sets of casework samples, and the SPC trends of data were monitored and used to adjust ‘calls’ for low-level samples. Thus, SPC may
offer a continuous and data-driven process for the evaluation and interpretation of low-level DNA peaks.
DNA, Interpretation, Statistical Process Control
26
Heteroplasmy, D-Loop, Mitochondrial DNA
B9
A Comparative Study of Y-STR Loci:
How do Different Sets of Y-STRs Fare
on a Common Population Panel?
Julie L. Maybruck, BA, BS*, The Ohio State University, 484 West 12th
Avenue, Columbus, Ohio 43210; Roger Kahn Kahn, PhD, Ohio Bureau
of Criminal Identification and Investigation, 1560 State Route 56
Southwest, PO Box 365, London, Ohio 43140; Paul A. Fuerst, PhD,
Department of Evolution, Ecology and Organismal Ecology and
Department of Molecular Genetics, The Ohio State University, 484
West 12th Avenue, Columbus, OH 43210
Via this presentation, attendees will be presented with the results of
a comparative analysis of several sets of Y-STR loci. A new set of YSTR loci developed using the human genome sequence is compared
with the most widely used Y-STR loci and with other recently identified
Y-chromosome loci.
A number of Y-STR loci have been identified and characterized;
however, there are concerns associated with some loci. We address these
concerns, identify new Y-STR loci, and perform a side-by-side examination of variability of new Y-STR loci with the most widely used YSTR markers and with a panel of other recently identified Y-STR loci.
Since it is important to identify which loci are the most appropriate and
informative for forensic applications, this study should have an impact
on the forensic community.
* Presenting Author
Proposition: Additional Y-chromosome microsatellite loci may be
needed for adequate forensic analysis for the following reasons: First,
the chromosomal distribution of currently available loci is mostly
limited to two small regions on the Y-chromosome; second, additional
loci may be needed because of potential typing errors caused by duplicated genetic material in the human genome; third, the loci may be
needed, as well, because of the relatively low level of variability of the
currently available unilocal loci.
Sex specific markers, such as Y-STRs, are highly valuable tools in
DNA forensics because men commit the majority of violent crimes.
Y-STRs can be used to distinguish the male component in body fluid
mixtures. They also aid in the identification of the number of male
contributors in multiple rape cases. Y-STRs are also useful in paternity
cases; particularly in situations involving a deceased putative father,
Y-STRs can identify patrilineage. In population studies, Y-STRs help to
identify paternal migration patterns, in contrast to maternal migration
patterns identified by mitochondrial DNA.
Six studies involving the identification/characterization of forensically useful Y-STRS are relevant: Kayser et al., 1997, White et al., 1999,
Ayub et al., 2000, Iida et al., 2001 and 2002, and Redd et al., 2002. In
terms of physical location within the Y-chromosome, loci in the first five
studies are mostly limited to two small regions fairly close to the Y-chromosome centromere. Several loci identified by Redd et al., 2002, are
also located in these same regions of concentrated loci. In addition to
the distribution within the Y-chromosome, a number of loci have additional drawbacks. Examination of information from the human genome
project and from the literature indicates that many current loci are duplicated elsewhere on the Y-chromosome or on the X-chromosome. Some
of these loci may be highly variable but, for forensics, are less than ideal.
Potential problems can occur during interpretation of genotype results.
For many forensic applications, the purpose of using Y-STR loci is to
preferentially amplify the male DNA contribution in mixed samples and
to determine the number of male contributors in multiple rape cases.
Duplicated loci defeat these purposes.
Following the identification of loci by the first three studies,
various combinations of markers have been used to examine populations, revealing a high number of unique haplotypes within populations.
The most widely-used Y-STRs are those of Kayser et al. Given potential
drawbacks of some available loci, we have identified new Y-STR loci.
About 26 Mb of Y-chromosome DNA sequence has been annotated by
the National Center for Biotechnology Information (NCBI). We
screened over 17 million bases of Y-chromosome sequence outside the
two concentrated regions of existing loci, identifying a library of 465
loci. BLAST searches against the human genome identified those that
were unique to one Y-chromosome location. Of 229 loci examined, 63
were determined to be unique to single Y-chromosome locations. The 63
loci were screened in a racially diverse panel of 30 individuals to ensure
unique amplification and to determine locus variability levels. Nearly
half were found to be highly polymorphic. The loci were further tested
for non-amplification in females. As a result, we chose a set of 10 male
specific marker loci dispersed across the Y-chromosome. We found 30
unique haplotypes in the 30 individual test population. Two Y-plex kits
exist which, together, contain the most frequently used loci. We
examined the same 30 individuals with these kits. Several haplotypes
occurred multiple times. The OSU 10-locus set had an average of 3 more
alleles per locus than the 10-locus Y-plex set. Since we identified over
60 loci by screening the human genome sequence, we expected to find
loci in common with other researchers. Examination of the loci identified by Redd et al. 2002, show 7 loci in common between our 63 loci
and the 14 Redd et al. loci. The OSU 10-locus set has an average of 2.5
more alleles per locus than the Redd et al. 7-locus set. (Note that only
one locus is common to both the Redd et al. 7-locus set and the OSU 10locus set). Further comparative analyses of the different sets of loci will
be presented.
A number of Y-STR loci have been identified and characterized;
however, there are concerns associated with some loci. We address these
* Presenting Author
concerns, identify new Y-STR loci, and perform a side-by-side examination of variability of new Y-STR loci with the most widely-used YSTR markers and with a panel of other recently identified Y-STR loci.
Since it is important to identify which loci are the most appropriate and
informative for forensic applications, this study should have an impact
on forensic science.
Y-STRs, Y Microsatellites, Y-Chromosome
B10
Forensic Analysis of Explosive
Residue Background
Steven H. Wise*, Olivier Collin, BS, John A. Mathis, MS, and Bruce R.
McCord, PhD, Ohio University, 136 Clippinger Laboratories, Athens,
OH 45701
methods for the detection of background levels of high explosives.
This presentation will establish parameters for detection of traces
amounts of explosives based on a nationwide, background-levels survey,
which will help investigators in eliminating false positives.
The ability to detect trace amounts of explosives is important in
forensic investigations. Various analytical techniques are effective for
the detection of explosives. In recent years, the sensitivity for the
detection of explosives has greatly increased. This often leaves the
forensic investigator in a situation where the significance of a result is
not just the detection of an explosive residue, but rather its concentration
when compared to that normally found in the environment. In order to
give the investigator the concentration threshold that minimizes false
positives, the background levels of explosive residues and interfering
compounds found in public places needs to be determined. The goal of
this project is to develop gas chromatography (GC) methods for trace
detection of organic residues of high explosives in environmental
samples.
This study involves the development of robust analysis methods for
trace amounts of the following compounds: ethylene glycol dinitrate
(EGDN), 4-nitrotoluene (4-NT), nitroglycerin (NG), 2,4-dinitrotoluene
(2,4-DNT), 2,6-dinitrotoluene (2,6-DNT), 2,4,6-Trinitrotoluene (TNT),
pentaerythritol tetranitrate (PETN), hexogen (RDX), octogen (HMX),
and tetryl. Sampling is done using store bought cotton swabs rinsed with
deionized-distilled water and isopropyl alcohol to remove impurities.
Samples are collected by a thorough swabbing of the area of interest
with dry, sterile cotton. Organic residues from each cotton swab are
removed by an acetone extraction followed by a volume reduction under
nitrogen flow. The extract is then screened for the presence of organic
explosives using an HP 6890 GC with electron capture detection. The
separation method involves a split-less injection at 180º C with helium
carrier gas and a ten-minute temperature program between 50 and
250º C. To reduce sample degradation during the separation, a megabore
capillary column is used; HP-5 (DB5 type, 95% dimethyl 5% diphenyl
polysiloxane; 10m length, 0.53mm diameter, 2.65µm film thickness).
This separation allows for presumptive identification and quantification
of each compound by external standards. Detection limits in the low
ng/ml range are possible for all compounds. The samples found to have
a positive screening result are then confirmed by GC-mass spectrometry
(MS) using negative chemical ionization with methane reagent gas. To
verify the presence of the selected explosives, GC-MS is performed
using a Finnigan GCQ equipped with a megabore Restek Rtx5MS (DB5
type, 15m, 0.53mm, 1.5µm film thickness) column that is split in the GC
by sliding the analytical column over a 0.1mm fused silica transfer line,
thus venting part of the flow into the oven. The risk of false positives is
reduced by the use of selective detectors and compound identification by
mass spectrometry. The analytical methods developed in this study will
allow for the creation of a database using samples collected across the
United States. To insure an accurate determination of background levels
in different environments, sampling will be done in many public places.
27
Such places include: malls, police stations, airports, taxis, and buses;
where door knobs, hand rails, counters, floors, walls, and furniture will
be sampled. The compilation of the results obtained from this study will
give the investigator a much needed tool for the analysis of organic
explosive residue in public places. In addition, this study addresses the
issues for the admissibility of explosives analysis results in court under
the Daubert ruling.
Explosives, Gas Chromatography, Mass Spectrometry
B11
Development and Validation of a
Y-Chromosome STR Typing System
Y-PLEX 12, for Forensic Casework
Jaiprakash G. Shewale, PhD*, Huma Nasir, BS, Elaine Schneida, and
Sudhir K. Sinha, PhD, ReliaGene Technologies, Inc., 5525 Mounes
Street, Suite 101, New Orleans, LA 70123
This presentation will demonstrate to the forensic community the
usefulness of Y-STR analysis in forensic casework.
The forensic community will have a better understanding of the usefulness of Y-STRs. The commercial Y-STR kit, Y-PLEX, can help identify
a Y-STR profile. Y-STR’s can benefit forensic casework and solve even
the most difficult sexual assault cases.
Short tandem repeat loci on the Y-chromosome (Y-STRs) have
become beneficial in resolving difficult forensic cases such as a sexual
assault case. Using Y-STRs, it is possible to obtain an exclusive profile of
male DNA in a sample containing mixtures of male and female DNA.
Scientific Working Group on DNA Analysis Methods (SWGDAM) has
identified a set of eleven loci namely DYS19, DYS385a/b, DYS389I,
DYS389II, DYS390, DYS391, DYS392, DYS393, DYS438 and DYS439
for forensic analysis (1).
Two Y-STR typing systems, Y-PLEX™ 6 and Y-PLEX™ 5, for
forensic DNA analysis are available commercially (2,3). The two systems
working together enable analysis for all 11 Y-STR loci recommended by
the SWGDAM. In order to achieve simultaneous amplification and
analysis for the 11 Y-STR loci, we have developed a Y-STR typing system
Y-PLEX™12. In addition to 11 Y-STR loci, the sex determinant marker
Amelogenin is incorporated in the Y-PLEX™ 12 system. Amelogenin provides results for gender identification and serves as internal control for
detection of PCR inhibitors in male/female mixture samples. The validation studies were performed according to the DNA Advisory Board’s
(DAB) Quality Assurance Standards. The minimal sensitivity of the YPLEX™12 system was 0.125 ng of male DNA. Amplification of DNA
from male primates, domestic and farm animals and microorganisms
reveal that the primers present in the Y-PLEX™12 system are specific for
human male DNA and some higher male primates. Female DNA, as high
as 700 ng, did not provide amplification products for Y-STRs. A database
for the 11 Y-STR loci for Caucasian, African American and Hispanic population groups, which is currently available at www.reliagene.com can
be used for obtaining haplotype frequency. The results reveal that YPLEX™12 is a sensitive, valid and robust multiplex system for forensic
analysis. Forensic casework examples demonstrating advantages of YSTRs will be presented.
1. Budowle B, Sinha SK, Lee HS, Chakraborty R. Utility of Y-chromosome STR haplotypes in forensic applications. Forensic Sci Rev
2003; 15: 153-64.
2. Sinha SK, Budowle B, Arcot SA, Richey SL, Chakraborty R,
Jones MD, et.al. Development and validation of a multiplexed
Y-chromosome STR genotyping system, Y-PLEXTM6, for forensic
casework. J Forensic Sci 2003; 48: 93-103.
3. Sinha SK, Nasir H, Gross AM, Budowle B, Shewale JG: Development
and validation of the Y-PLEXTM5, a multiplexed Y-chromosome STR genotyping system, for forensic casework. J Forensic Sci 2003; 48 (5): In Press.
Forensic Casework, Y-STR, Y-PLEX
28
B12
Trace Element Profiles of Float Glass
Fragments Determined by Laser
Ablation Inductively Coupled Plasma
Mass Spectrometry (LA-ICP-MS)
David W. Szymanski, BA, MS* and Lina Patino, PhD, Department of
Geological Sciences, Michigan State University, East Lansing, MI
48824; Christopher Bommarito, MS, Michigan State Police, Forensic
Science Division, Lansing, MI 48913; Jay A. Siegel, PhD, School of
Criminal Justice, Michigan State University, East Lansing, MI 48824
The goal of this presentation is to present a method for forensic glass
analysis by LA-ICP-MS for comparison and standardization of this new
application.
LA-ICP-MS in forensic science is highly promising new technique for
the rapid discrimination of glass fragments by elemental composition. The
relatively quick, non-destructive nature of LA, coupled with the sensitivity
of ICP-MS makes the method well suited for glass analyses. Development
of a protocol for the equipment in this study sets the groundwork for implementing a standard inter-laboratory technique. The results from this study
can be compared with those from other laboratories, aiding in the standardization of the method and validating the technique for casework.
The importance of glass as trace evidence, coupled with increasing
physical and chemical homogeneity of float glass, has prompted the use of
innovative techniques for discriminating between samples. Inductively
coupled plasma mass spectrometry (ICP-MS) is becoming a widely used
method for elemental analyses of glass fragments, with laser ablation (LA)
of solid samples being the most efficient introduction system for forensic
applications (Trejos et al., 2003; Watling et al., 1997). LA-ICP-MS
requires almost no sample preparation and consumes extremely small
amounts of sample, making it a virtually non-destructive technique.
This poster will outline an instrumental protocol for forensic glass
analysis developed at the ICP-MS Laboratory at Michigan State University.
Using a Cetac LSX 200 Plus Nd:YAG laser (266 nm) attached to a
Micromass Platform quadrupole ICP-MS, a certified standard (NIST 612)
and six unknown automobile float glass fragments provided by the
Michigan State Police were used to optimize ablation parameters for
acquiring eight trace elements (85Rb, 88Sr, 89Y, 90Zr, 98Mo, 138Ba, 139La,
and 140Ce). The technique was optimized for the smallest spot size (best
for forensic applications) that produced adequate signal above background
for all isotopes. For both the standard NIST 612 and the unknown, these
conditions were met with a spot size of 100 µm and depth profile z-rate of
1 µm/s for 30 s, fixing the crater depth at 30 µm. The detector was initiated
before ablation and recorded for a total of 60 s. For each trial, a 1 s preablation burst of the laser was focused at the sample surface to ensure it was
free of contaminants. Both the float and non-float sides of the unknown
glass fragments were ablated five times to test the homogeneity of the
samples and precision of the technique with this experimental setup.
Similarly, a cross-sectional analysis of one fragment was conducted using
the same parameters, with spot analyses 300 um apart, to ensure homogeneity and precision in the case of fragments without parallel sides.
Data were reduced by a standard gaussian integration of peaks,
obtaining the maximum peak height above background for each element.
The precision of the technique was evaluated by element using normalized
peak heights. Overall, the precision of replicate analyses is very good
(<10% RSD), within and between replicates on both the float and non-float
sides of samples. Only two elements, Mo and Rb, which had the lowest
peak heights, consistently show variation >10% RSD. The results of the
experiments can be presented graphically on triangular plots of element
ratios after Watling et al. (1997), demonstrating the discriminating power of
the method. Using only six elements (88Sr, 89Y, 90Zr, 138Ba, 139La, and
140Ce) from the analyses, out of a total of 15 unique pairs, all glasses were
distinguishable by this method.
Laser Ablation ICP-MS, Glass Fragments, Trace Elements
* Presenting Author
B13
Trace DNA: Casework Experience
Carolyn L. Booker, BS, George J. Schiro, Jr., MS*, Winnie C.
Wong, MSc, and Ray A. Wickenheiser, BSc, Acadiana Criminalistics
Laboratory, 5004 W Admiral Doyle Drive, New Iberia, LA 70560
The learning objective of this presentation is to familiarize forensic
scientists with the concept and practical applications of trace DNA
analysis.
This presentation adds to the developing trace DNA casework
analysis body of knowledge. For those in the forensic science community not familiar with the advantages and disadvantages of trace DNA
analysis, this presentation will introduce its conceptual and practical
applications. Forensic scientists who routinely conduct trace DNA
analysis can compare their findings to the data in this study. As this data
accumulates, the best methods for analyzing trace DNA can be
developed.
Trace DNA is defined as the minute quantities of DNA transferred
through skin contact, which can be successfully analyzed and follow the
general principles of trace evidence. Polymerase chain reaction (PCR)
technology has made the analysis of short tandem repeats (STRs) possible on the most minute and degraded DNA samples, such as trace DNA
samples. Trace DNA STR analysis is a relatively new field in forensic
science. Similar to the general principles of trace evidence, this type of
analysis involves analyzing areas of potential skin contact to determine
if an STR DNA profile can be obtained from these areas.
At the time of this publication, the Acadiana Criminalistics
Laboratory (ACL) had analyzed 105 potential DNA trace samples over
an 11-month period. Typical samples analyzed include steering wheels,
cloth and latex gloves, caps/hats, tools, firearms, clothing, latent print
smudges, and commercial containers. Using previously validated STR
analysis procedures and adhering to the ACL’s STR interpretation guidelines, the lab was successful in obtaining DNA profiles from 71% of the
samples. 34% of the samples had results at 14 loci (13 STR CODIS core
loci and amelogenin) and 37% of the samples produced partial profiles
of 13 or fewer loci. 29% of the samples produced no results. Of the
samples that produced DNA profiles, 55% were mixtures, 41% appear to
have originated from a single source, 3% of the profiles were not interpretable, and 1% was traced to a contaminating source. When compared
to reference samples submitted for comparison to the potential trace
DNA samples, the reference samples were excluded as the source of the
DNA in 31% of the samples. The reference samples were included as a
possible source of the DNA in 44% of the samples. No conclusion as to
the source of origin of the DNA could be drawn in 23% of the samples.
1% of the samples had contamination that could be traced to a known
source.
Specific collection techniques, analytical methods, STR interpretation guidelines, updated information, and case examples will be presented at the meeting.
Trace DNA, STRs, PCR
B14
Increasing the Efficiency of STR
Profiles Through Amplified Product
Concentration
Gina M. Sola, BS, MFS*, Susan E. Welti, BA, MFS*, James M.
DiFrancesco, MFS, Timothy P. McMahon, PhD, Theodore D.
Anderson, MFS, Demris A. Lee, MSFS, and Brion C. Smith, DDS,
Armed Forces DNA Identification Laboratory, 1413 Research
Boulevard, Building 101, Rockville, MD 20850
humanity by providing an alternative to re-amplification which preserves sample extract and increases the possibility of generating a profile
* Presenting Author
in the absence of any remaining extract.
In forensic nuclear DNA analysis, DNA profiles for unknown case
samples are generated using one of several commercially available short
tandem repeat (STR) kits. A common occurrence with inhibited or
degraded samples is the generation of a partial profile (low-intensity
alleles), or no profile at all. However, there may not be enough extract
or specimen remaining to re-amplify these samples. The goals of the
experiments described in this presentation were to determine if the concentration of amplified STR products by vacuum evaporation could
increase the relative fluorescence units (RFUs) of the alleles that were
originally below the reporting threshold, and if so, to validate this concentration method for use at the Armed Forces DNA Identification
Laboratory (AFDIL).
In order to validate the concentration procedure, 6 samples of 1.0
ng positive control DNA were amplified and analyzed with the ABI
AmpFlSTR® Profiler Plus and AmpFlSTR® Cofiler kits on the ABI
377. All of the alleles from these samples were over AFDIL’s 50 RFU
reporting threshold. One of the positive DNA control amplification
products from each kit was serially diluted as follows: (a) 1:4, (b) 1:8,
(c) 1:16, (d) 1:32, (e) 1:16, (f) 1:32, (g) 1:64, and the dilutions were analyzed on the ABI 377. Dilutions that generated either no STR results or
partial STR profiles (profiles that had allele peaks below 50 RFUs) were
used as the standard by which nine of the remaining ten STR amplification products were diluted. Results demonstrated that 120 out of 150
possible allele peaks were below 50 RFUs for Profiler Plus and 104 out
of 120 possible allele peaks were below 50 RFUs for Cofiler.
To test whether concentration of the diluted product would increase
the allele peak RFUs, the diluted amplification products (described
above) were transferred to 1.7 mL tubes and dried down completely in a
Jouan HetoVac for 1 hour. The concentrated amplification products
were then re-suspended in a loading solution containing 4 uL distilled
water and 5 uL formamide loading buffer (formamide, GS500, and
Bromophenol blue) and reanalyzed. The resulting STR profiles were
compared to their corresponding diluted profiles for increases in peak
RFUs. On average, the peak heights for Profiler Plus increased 4-fold
and for Cofiler 6-fold. After concentration, 116 out of the 120 Profiler
Plus peaks previously below AFDIL’s 50 RFU cut off were now above
the reporting threshold. Likewise, 96 out of 104 allele peaks that were
below reporting thresholds for Cofiler were now above 50 RFUs. In no
instance were additional extraneous peaks observed for any sample after
concentration.
The concentration method was then applied to non-probative
casework reagent blanks, negatives, and substrate controls to determine
if any minor contaminates that were not detectable prior to concentration
were evident after concentration. A total of 39 controls consisting of 16
Profiler Plus negatives and 19 Profiler Plus reagent blanks and 1 Cofiler
negative, 1 Cofiler reagent blank, and 2 Cofiler substrate controls that
had no visible peaks prior to concentration were concentrated. After
concentration, 32 of the 39 controls had no visible peaks; however, the
remaining seven controls exhibited 1 or 2 peaks that Genotyper called as
true alleles. The concentrated profiles were then compared to the
original sample profiles from their respective cases and in no instance
did the concentrated peaks match any of the original case sample profiles, suggesting that these peaks were most likely the result of concentrating a low-level PCR contaminant that was present in the unconcentrated sample.
As the final validation step, twenty-two casework samples from
Qiagen BioRobot extracted bloodstain cards that had exhibited partial
Profiler Plus profiles were concentrated and analyzed as described
above. Results demonstrated that 80% of the peaks, or 74 out of 92 possible allele peaks increased in RFUs after concentration. Of the 74 peaks
that increased in RFUs, 52% or 48 peaks were brought above AFDIL’s
50 RFU-reporting threshold. The average peak height increased 3.5fold. In all instances but one, the post-concentration peaks were in concordance with the original STR profiles generated during case pro-
29
cessing. In one sample, an extraneous peak was observed at the FGA
locus. This peak, however, was not present after the sample was
reloaded, arguing that the peak was most likely due to a gel artifact.
In conclusion, concentration of STR amplification products has
proven to be effective in improving the chance of obtaining an STR
profile when no profile or a partial profile was originally generated after
amplification. Concentration provides an alternative to re-amplification,
which preserves the sample extract and increases the possibility of generating a profile in the absence of any remaining extract. In addition, the
baseline background for the samples is not elevated after the concentration process, which suggests that this process does not introduce artifacts into the samples that are not already present at low levels.
However, it is recommended that all negatives, reagent blanks, and substrate controls be concentrated along with their corresponding casework
samples for comparison on a case-by-case basis.
The opinions and assertions expressed herein are solely those of the
authors and are not to be construed as official or as the views of the U.S.
Department of Defense or the U.S. Department of the Army.
STR, Partial Profile, PCR Product Concentration
B15
Potential Contamination When Wearing
Sterile Gloves During PCR Preparation:
Pass-Through Contamination From Skin
Miguel Lorente, MD, PhD*, Institute of Legal Medicine, University of
Granada, Av. Madrid 11, Granada, 18012, Spain
Contamination during DNA analysis based on PCR is a serious
concern that usually happen in genetic labs. Although protocols are
developed to avoid this issue, it still happen and most of the cases is not
possible to find out the source of contamination. This paper present how,
even wearing sterile gloves, contamination from the user can happen. It
means that some extra-means should be considered in this sense.
humanity by solving some issues regarding to contamination and crosscontamination, and, at the same time, providing knowledge and focus
attention to some circumstances that are not frequent, but when they
happen can affect the result of a case
1. Introduction: Mitochondrial DNA (mtDNA) analysis has became
a routine procedure in human identification and in anthropological
studies. One of the advantages of analyzing mtDNA is the enhanced sensitivity afforded with the technique. But this feature must be considered
because contamination can affect the final results of a study. Quality
control and quality assurance procedures are enacted to minimize and
monitor contamination. However, sometimes it is not easy to identify the
source of spurious or inconsistent. One vector for contamination is the
gloves worn during experimentation. It is imperative to wear sterile
gloves and change the gloves as needed. Clinical and epidemiological
studies have demonstrated that bacterial and viral contamination can
occur on the surface of sterile gloves after being worn for a length of
time. Thus, DNA may get on gloves and be transferred during handling
(i.e., cross contamination). It also is possible that if gloves are worn for
certain periods of time that DNA may leach from the user’s hand through
the glove (i.e., pass through contamination). While contamination of this
nature is not a routine concern, it may explain rare circumstances of
undefined contamination. Therefore, we designed a set of experiments to
determine if gloves could be conduits of contamination.
2. Material and Methods: To study pass-through contamination
gloves were worn for different time periods compatible with labs tasks.
Gloves were worn without touching anything for 5, 10 and 20 minutes
by different users. Only intermittent gentle rubbing between the thumb
and index finger was carried out to mimic general manipulations.
After each time frame a sample was taken from the areas usually in
contact with the tubes using a wet cotton swab and a negative control
30
was taken from a zone where no manipulations occur. After the
swabbing the gloves were discarded.
All the swabs were extracted using an organic method (PCIA) and
amplified for HVIB and HVIIA according to Wilson et al., we also
included some nuclear DNA amplification. Post-amplification of the
nuclear and mtDNA product was carried using capillary electrophoresis
as previously described.
3. Results and Discussion: The experiments show that the length of
time gloves are worn is an interesting factor to be considered. In some
samples even after five minutes some DNA leached through the gloves,
even in the apparent negative controls. These results are compatible with
the clinical studies which have shown that after a time, even with careful
washing, bacterial and virus contamination on the surface of the gloves
can occur related to time and user.
These findings do not suggest that new practices in contamination
control are warranted. They do suggest possible sources for contamination when it occurs. If gloves are not changed between cases, cross
contamination may occur and explain why DNA types from unknown
sources may be observed. However, cross contamination is not a serious
concern under current practices. Pass through contamination may
explain the presence of the operators mtDNA in a sample. Sensitivity of
mtDNA analysis requires special care during the handling of samples
and reagents, and particularly in extreme situations where sample
manipulation is for prolonged times. If contamination persists, one may
consider changing gloves every five-ten minutes or using double gloves.
Also, washing the hands prior to putting on gloves could remove dead
cells or their products from skin surface.
DNA, Contamination, Gloves
B16
The Effect of Formalin Decontamination
on STR Analysis Conducted on Human
Remains Submitted for Identification
Deborah K. Haller, BS*, Armed Forces DNA Identification Laboratory,
1413 Research Boulevard, Building 101, 2nd Floor, Rockville, MD
20850-3125; Susan W. Jones, PhD, Demris A. Lee, MSFS, Timothy P.
McMahon, PhD, and Brion C. Smith, DDS, Armed Forces DNA
Identification Laboratory, 1413 Research Boulevard, Rockville, MD
20850; Craig Mallak, MD, Armed Forces Medical Examiner System,
1413 Research Boulevard, Building 102, Rockville, MD 20850
After attending this presentation, attendees will be familiar with the
use of 10% formalin in decontaminating human remains and the effect
of the decontamination on Short Tandem Repeat (STR) DNA analysis of
bone and tissue specimens.
This presentation will show the forensic community that storage of
human remains in 10% formalin for up to 30 days will have little to no
effect on the ability to obtain an STR profile.
The mission of the Armed Forces DNA Identification Laboratory
(AFDIL) is to assist the Armed Forces Medical Examiner System
(AFMES) in identifying members of the armed services using nuclear
DNA methods. The recent war in Iraq raised the specter that biological
and/or chemical weapons could be used against American troops. A
decontamination scheme had to be developed so that DNA testing could
be used to identify fallen service men while ensuring the protection of
all people having contact with the biologically contaminated remains.
One of the methods examined to potentially decontaminate the human
remains was storing human remains in 10% formalin for a period of
time. 10% formalin is known to kill biological agents such as anthrax.
However, it is also known that formalin can have a detrimental affect on
the DNA contained within the sample. To determine if it was possible
to obtain a Short Tandem Repeat (STR) profile from biological samples
that had been stored in 10% formalin, the middle portion of an index
finger or the tip of an index finger from medical cadavers were sub* Presenting Author
mersed in 10% formalin for 5 days, 7 days, 10 days, 15 days, and 30
days. The samples were then removed from the 10% formalin, the tissue
was dissected away and stored at –20 ºC, and 0.9 grams – 2.0 grams of
bone was immediately extracted using the AFDIL’s organic protocol for
extracting DNA from bone samples. Quantitation of the samples determined that approximately 100 nanograms of DNA per microliter was
recovered. The samples were amplified using the AmpFlSTR® Profiler
PlusTM Amplification and Typing Kit. The amplicons were run on an
ABI Prism 377 and analyzed using Genotyper version 2.5. Likewise, the
tissue samples were extracted following the AFDIL’s organic protocol
for extracting DNA from tissue samples. The amount of tissue extracted
was approximately 5 mm x 9 mm in size. These samples were done in
triplicate with the only difference between the extraction sets being a
wash step prior to extraction. One set of extractions was conducted as
per the protocol (i.e. no wash step). A second set of extractions was conducted after a portion of the tissue had been washed with sterile distilled
water. The third set of extractions was conducted after a portion of the
tissue was washed with 150mM glycine. The extracted DNA from the
tissue samples was amplified and typed as described above. Full STR
profiles were obtained in nine (samples stored for 5 days, seven days,
and ten days) of the eleven bones extracted. The sample submerged in
formalin for 15 days gave results at all loci except D18S51 and D7. The
sample submerged in formalin for 30 days gave results at all loci except
D7S820. The profiles obtained matched the “expected profiles” and
there were no mutations observed in the samples that had been stored in
the formalin. It is possible to obtain nuclear DNA profiles from specimens stored in formalin up to 30 days.
authors and should not be construed as official or as views of the United
States Department of Defense or the United States Department of the
Army.
1Armed Forces DNA Identification Laboratory
2Armed Forces Medical Examiner System
Human Remains, STR Profile, Biological Weapons
B17
Laser Ablation/Pulsed-Field Reflection
TOF-MS Analysis and Discrimination
of Forensic Soil
Sung-Woo Park, PhD, Eun-Ho Kim, MS*, National Institute of
Scientific Investigation, 331-1 Shinweol-7-Dong, Yangcheon-ku, Seoul,
158-707, South Korea; Il-Kwang Kim, PhD, Department of Chemistry,
Wonkwang University, 344-2 Shinyong-Dong, Iksan, Jeonbuk, 1,
Korea; Kwang-Woo Jung, PhD, Department of Chemistry, Wonkwang
University, 344-2 Shinyong-Dong, Iksan, Jeonbuk, 1, Korea
After attending this presentation, attendees will learn how to
analyze and discriminate forensic soil evidence using Laser
Ablation/Pulsed-field Reflection TOF-MS.
This presentation will introduce a new method of analyzing and
discrimination of soil samples by using Laser Ablation/Pulsed-field
Reflection TOF-MS.
This presentation will discuss how to analyze and discriminate
forensic soil evidence using Laser Ablation/Pulsed-field Reflection
TOF-MS.
Soils vary with areas and have distinct characteristics due to natural
effects and the residues of leaves and living things. Due to complex
variations in compositions, several examination techniques and instruments should be used for analyzing soil samples. In most cases, physical
examinations of color reaction, a polarization microscope, etc and
instrumental analyses of XRF and SEM-EDX have been used to discriminate forensic soil samples.
In this study to discriminate soil evidences, 30 soil samples were
collected from 30 different locations in Republic of Korea and were analyzed using Laser Ablation/Pulsed-field Reflection TOF-MS.
* Presenting Author
All samples were reduced to powders and then were made into
pellets (ID 0.8mm). The samples were heated for 5 hours at 500º C
before loading on the holder. The Nd:YAG laser light (^=532 nm) was
used to ablate the components of soil. The mass spectrum was obtained
by averaging of repeated pulses of 2000 times checked by Oscilloscope.
The ablated main elements were sodium (Na), aluminum (Al),
potassium (K), titanium (Ti), iron (Fe), copper (Cu) on the Mass
spectrum. The ratios, (Al)/(Ti) and (Fe)/(Ti), showed reproducibility and
different results with the soils prepared from the different locales. It
could confirm that these results can be used for forensic soil examinations and supported to other analyses: data obtained by a polarization
microscope and SEM-EDX, and so on. The isotope ratio for Titanium
(the ratio of 45.95amu to 46.95amu) also showed another useful criterion to discriminate soil samples. This data gave the same results as the
ones analyzed by the ratios of components.
Thus, these could be useful techniques to discriminate forensic soil
samples by comparing both the component ratios, (Al)/(Ti) and
(Fe)/(Ti), and Isotope ratio of Ti. Applications were also carried out on
forensic soil evidence submitted from the police’s. It showed much more
definite results compared by the ones obtained by routine analyses with
a polarization microscope and SEM-EDX.
It was concluded that analyzing soil evidence by Laser
Ablation/Pulsed-field Reflection TOF-MS could be a useful technique to
identify forensic soil evidence.
Forensic Soil Evidence, Laser Ablation/Pulsed-Field Reflection
TOF-MS, Mass Spectrum
B18
The Design and Compilation of a
Comprehensive U.S. Y-STR
Haplotype Reference Database
Paulina N. Berdos, BS* and Erin K. Hanson, MS, National Center for
Forensic Science, University of Central Florida, Department of
Chemistry, PO Box 162367, Orlando, FL 32816-2367; Stacey Smith,
MS, South Dakota DCI Forensic Laboratory, 3500 E Highway 34,
Pierre, SD 57501; Jack Ballantyne, PhD, National Center for Forensic
Science, University of Central Florida, Department of Chemistry, PO
Box 162367, Orlando, FL 32816-2367
After attending this presentation, the forensic community will learn
the design, establishment and statistical analysis of a comprehensive online Y-STR Database.
This project would impact the forensic community because it will
provide reliable estimates of Y-STR haplotype frequencies and provide
a statistical estimate of the significance of a match.
The establishment of a U.S. National Y-STR reference database
from a variety of geographically and ethnically diverse populations is
essential to facilitate the generation of reliable estimates of Y-STR haplotype frequencies. Such multi-locus haplotype frequencies are required
to provide a statistical estimate of the significance of a match. A U.S. YSTR Haplotype Reference Database has been created by the
International Forensic Y-User Group and is maintained by the Max
Planck Institute for Evolutionary Anthropology, Leipzig, Germany.
However, this database has been limited to a set of 9 core Y-STRs that
limits its operational usefulness, particularly in light of the development
of Y-STR multiplexes consisting of additional Y-STR loci. Y-STR loci,
unlike traditional STR markers, are not independent of one another and
are co-inherited as extended haplotypes of linked markers. The estimation of the frequency of occurrence of a particular haplotype therefore
necessitates the use of a counting method which means that the significance of many matches are dependent upon the size of the database.
The NCFS database initially comprised data generated in the laboratory based upon a 19 Y-STR locus extended haplotype. The loci
include DYS19, DYS385 (a) and (b), DYS388, DYS389I, DYS389II,
DYS390, DYS391, DYS392, DYS393, DYS425, DYS434, DYS437,
31
DYS438, DYS439, Y-GATA C4, Y-GATA A7.1, Y-GATA A7.2 and YGATA H4. Data was obtained from various Caucasian, African
American, Hispanic, Native American and Asian populations. A comprehensive evaluation of the data was carried out to determine the allelic
distribution and the gene diversity at each locus as well as the multilocus discriminatory potential. Allelic ladders for the 19 Y-STR markers
have been constructed to aid in accurate genotype determinations. Since
there are a number of common 19-locus haplotypes additional loci have
been included in the database. These loci include DYS426, DYS436,
DYS441, DYS450, DYS462, YAP (Alu insertion), Y-GATA-A10,
DYS531, DYS557 and DYS588. Although, some data exists for some of
these loci in U.S. populations, it is not readily accessible to the crime
laboratory community and usually does not contain individual multilocus haplotype data.
A key component of our strategy is to allow for the continuous
updating of haplotype data using the same samples. This ensures that as
new markers are developed, the same samples would be re-typed, and a
new extended haplotype developed. Thus, any laboratory needing haplotype data for any combination of Y-STR markers would be served. The
aid of geographically diverse crime laboratories has been enlisted to obtain
the necessary samples. In exchange for the samples the crime laboratories
benefit by obtaining a custom built no-cost local Y-STR database.
Our results to date will be presented, and information on community access will be provided.
Y-STR Database, Haplotype Reference Database,
Populations Studies
B19
Guidelines for the Interpretation
and Reporting of STR Profiles:
A Hypothesis-Driven Approach
Alex Welsh, MSc* and Tara Brutzki, MSc, Centre of Forensic
Sciences, 25 Grosvenor Street, Toronto, ON M7A 2G8, Canada;
Tania Burrows, BSc, Centre of Forensic Sciences, 70 Foster Drive,
Sault Ste Marie, ON P6A 6V3, Canada; Silvana Tridico, BSc and
Jack Laird, MSc, Centre of Forensic Sciences, 25 Grosvenor Street,
Toronto, ON M7A 2G8, Canada
After attending this presentation, participants will learn about the
adoption of a hypothesis-based testing model for the interpretation and
reporting of DNA STR profiles.
We consider it vital to carry out all forensic biology casework
processes, including the interpretation and reporting of STR profiles, in
consideration of the hypothesis being tested. We will show how we have
adapted our guidelines to take various elements of the case history into
account in the interpretation and reporting stages of the analysis, and how
these in turn lead to clearer reports.
This poster will describe the guidelines used at the Centre of
Forensic Sciences for the interpretation and reporting of STR profiles,
recently revised and written within the framework of a hypothesis-based
approach to forensic casework. This approach can be summarized as a
mechanism to test hypotheses, determine if the results support such proposals and to report the conclusions in a manner reflective of the
hypotheses being tested.
It is the role of the scientist to address the pertinent questions being
asked, relevant to the events recorded in the case history, and to then make
a decision as to whether the testing of particular items would allow one to
draw valid inferences in relation to the hypothesis. The assumptions that
can be made during an interpretation may be based on both the DNA
results themselves and/or on elements of the case history. Scientists
however must also be prepared to address their conclusions under a different hypothesis with a different set of assumptions.
32
Examples of the interpretation process will be provided, along with
respective supporting rationales. Illustrations of the following steps in the
process will be provided:
(1) Deriving constituent DNA profiles from mixtures.
Dual source mixture: one source known or assumed.
Dual source mixture: both sources unknown.
Dual source mixture: equivalent major contributions of 2
unknown sources.
(2) Determining if a DNA profile is suitable for comparison purposes.
When making this determination, factors to consider individually or together include the amount of DNA detected, a lack of peak
height concordance, a prior expectation of background DNA (e.g. on
clothing) and the total number of contributors. If the determination is made
that the DNA profile is not suitable for comparison, the reporting of this
finding should reflect the reasons why this is so, for example:
“Due to uncertainty with respect to the number of contributors and/or due
to the low amounts of DNA detected, this DNA profile is not suitable for
comparison.”
(3) Reporting of results when a DNA profile is suitable for
comparison.
Once a DNA profile is determined to be suitable for comparison, one of two different formats of reporting will be required
depending on whether comparison samples from known individuals are
available.
(i) Can an individual in question be excluded as the source? The significance of an individual not being excluded as a source is routinely
expressed in the form of a random match probability (RMP) statement
(excepting results from familial analyses). For example:
“Mr. X cannot be excluded, at 9 STR loci, as the contributor of the
DNA profile from item 1. The probability that a randomly selected individual unrelated to X would coincidentally share the observed DNA profile
is estimated to be 1 in Y.”
(ii) If no known comparisons are available the DNA profile’s suitability for comparison at a later date can be reported in a number of ways
depending on its rarity:
“This DNA profile is suitable for comparison.”
“This DNA profile is suitable for comparison, though it may be of
limited forensic significance due to its relatively high* frequency of occurrence. * This DNA profile is expected to occur in the population with a frequency of greater than 1 in 1000.”
(4) Assessing the forensic significance of a DNA profile or of one
or more constituents in a mixture.
A determination is made as to whether the interpretation of each constituent profile in a mixture is necessary, based on the sample type and case
history. Reporting of results are worded to reflect this determination.
Examples will be given for the application of this assessment as relevant to:
Intimate samples – DNA analysis of an intimate sample (defined as
swabs from body orifices, skin swabs, fingernail scrapings and underwear
in certain circumstances) will generally be expected to yield the DNA
profile of the donor, in addition to DNA from someone else. The presence
of DNA from the person from whom the sample was taken is not of
forensic significance (unless the source of the sample is itself questioned).
However, given the prior expectation of detecting the donor’s DNA, this
knowledge can be used in the derivation of any additional contributing
DNA in the mixture. Examples of wording such findings include:
“No DNA, other than that which is attributable to Ms X, was
detected on a swab of her skin.”
“In addition to DNA attributable to Ms X, a male DNA profile was
determined at 9 STR loci from a swab of her skin and is suitable for comparison.”
Clothing – it is not unusual to find detectable levels of DNA on items
of clothing and as a consequence clothing can often be examined to
determine if it can be associated to a particular individual at some point in
time, in the absence of testing for a body fluid. However, as clothing is
often examined solely for the purpose of determining the source of a body
* Presenting Author
fluid stain there may or may not be significance to any additional DNA
coincidentally detected in the background. It should be noted nonetheless
that in some instances a minor level of background DNA might indeed be
of forensic significance, depending on the hypothesis being tested and the
elements of the case history. An example of wording to account for this
possibility would be:
“Mr. X cannot be excluded as the source of the bloodstain on the
jeans. [insert RMP statement]. An additional minor source of DNA was
also detected in the sample. Since it is not unusual to detect low levels of
DNA on clothing, this finding may be incidental and of no forensic significance.”
It is recognised that the development of guidelines for interpretation
is a continually evolving process requiring constant review and that the
scientific laboratory itself should play a role in determining how best to
provide the results to the clients in a forensically relevant and time efficient format.
Interpretation Guidelines, STR, Hypothesis Testing
B20
The Arab Population Data for 10
Y-Chromosome Specific STRs
Hisham E. Ragab, MD*, Ahmed H. Al-Awadi, PhD, Safia M.
Abdulla, BS, Lathqia A. Kassem, BS, and Nisreen W. Haddad, MS,
Abu Dhabi Police Forensic Science Laboratory, PO Box 32404,
Abu Dhabi, United Arab Emirates
After attending this presentation, attendees will understand the differences and similarities between the studied Arab Population groups
and other population groups.
This presentation will impact the forensic community by adding to
the Y-Chromosome population database as until now there is no
available Arab population data.
Objective: The objective of the present work: 1.to establish basic
data base as regards the 10 Y-chromosome STR loci for 6 Arab population groups, and 2. to evaluate the significance of an inclusion with Ychromosome specific STRs after establishing haplotype frequencies for
the examined population groups.
Nature of Study: Population Study.
Materials, Methods and Results: The study was conducted on
blood samples which were collected from unrelated healthy adult males
from 6 Arab population groups living in Abu Dhabi, United Arab
Emirates (U.A.E.) including native population in Abu Dhabi, and from
Egypt, Syria, Sudan, Jordon and Oman. DNA was extracted by both the
organic phenol-chloroform, and FTATM paper extraction protocols,
After quantitation of the extracted DNA (organic extraction only) amplification was carried out using about 2 ng of genomic DNA in a total
reaction volume of 25 µl - one amplifications - for 10 loci - (DYS19,
DYS385, DYS389I, DYS389II, DYS390, DYS391, DYS392, DYS393,
DYS438 and DYS439) using Y-STR kit from Promega. The amplified
product was tested on the ABI 310 genetic analyzer and the obtained
profiles were interpreted and analyzed using Gene Scan analysis
software (PE Applied Biosystems) and the genotypes were determined
by using the Genotyper DNA fragment analysis software (PE Applied
Biosystems) with the Power Typer Y Macro. Haplotype frequencies
were calculated for each STR locus for U.A.E. population and other population groups, frequencies were calculated through the gene counting
method, haplotypes and gene diversity was calculated. The obtained
results were compared with relevant Arab and other ethnic groups
database.
B21
Population Data on 9 STR
Loci in Crete (Greece)
Gabriel Petridis, BS* and Ersi Abaci-Kalfoglou, PhD, Istanbul
Unversity, Institute of Forensic Sciences Cerrahpasa, Istanbul, 34303,
Turkey; Maria Christakis-Hampsas, PhD and Yasmine Kapsali, MS,
Center of Toxicological Sciences and Research, University Hospital of
Heraklion, Medical School University, Heraklion, Crete 1515, Greece;
Sevil Atasoy, PhD, Istanbul Unversity, Institute of Forensic Sciences
Cerrahpasa, Istanbul 34303, Turkey; Manolis Michalodimitrakis, MD,
Department of Forensic Pathology, University Hospital of Heraklion,
Medical School University, Heraklion, Crete 1515, Greece
After attending this presentation the attendees will learn data on
CSFPO D7S820 D13S317 D16S539 F13A01 FESFPS THO1 TPOX
and vWA STRs in the population of Crete and their validity for the use
of forensic analysis.
This presentation will impact the forensic community by providing reliable STR data for probability calculations in cases where
Cretan population is involved
Crete is the largest of the Greek islands and is located in the South
East of the Mediterranean Sea. It is 260 km long and 60 km at its
widest point. Being an island, it is considered to be fairly isolated and
since geographically well separated, gene frequency differences were
expected. There is a lack of population genetic data from most of these
STRs despite the fact that the commercially available amplification
kits are increasingly being used in forensic casework. Therefore we
aimed to evaluate the data for the above-mentioned polymorphic loci,
for an effective use in Crete and the suitability of the system for
forensic applications.
Four hundred samples from unrelated individuals were tested for
the systems as for their gene and phenotype frequencies. The sampling
was done considering the population distribution in the four main residential areas of the island.
Following the DNA extraction and quantitation, the PCR amplification of the nine STR loci (CSFPO D7S820 D13 S317 D16S539
F13BA01 FESFPS THO1 TPOX and vWA) was performed using the
commercially available triplex amplification kits provided by
Promega, and the application was done according to the manufacturer’s instructions.
Allele frequencies were estimated by gene counting. The expected
genotype frequencies were calculated on the basis of estimated allele
frequencies under the assumption of Hardy-Weinberg equilibrium.
Accordance with Hardy-Weinberg equilibrium was checked using the
software package Popgen32. Observed and expected heterozygosity,
power of exclusion, power of discrimination, probability of match and
paternity index was also calculated.
The observed genotype distributions of the nine STRs showed no
deviations from Hardy-Weinberg expectations. The obtained results
were compared to the present data for Greek population from different
geographical areas and to the data for the rest of the Europe. No significant differences were observed between the populations for any of
the STRs analysis.
In no cases the observed heterozygosity is less than that of
expected. The discriminatory power and exclusion probability values
for all the analyzed markers are significantly high and thus reveal high
forensic significance. This allele frequency data will be useful for
human identity testing in Cretan population.
STRs, Crete, Greece
Forensic DNA, Y-Chromosome STRs, Arab Population
* Presenting Author
33
B22
Development of a Y-STR Megaplex and
Its Comparison to A Y-STR 10-Plex
Cassie L. Johnson, MS*, Robert C. Giles, PhD, Joseph H. Warren, BA,
and Rick W. Staub, PhD, Orchid Cellmark, 2600 Stemmons Freeway,
Suite 133, Dallas, TX 75207
After attending this presentation, attendees learn about new Y-STR
multiplexes that are being developed for use in forensic and paternity
laboratories.
This presentation will demonstrate that Y-STRs are being used
increasingly more in forensic and paternity laboratories. The Y-STR
multiplexes being developed at Orchid are able to quite effectively discriminate among male lineages, and thus, will have a significant impact
in the field of human identification.
Y-chromosome STRs (Y-STRs) have gained interest in the forensic
community because of their ability to identify the male component of a
sample. Y-STR testing is especially valuable in sexual assault cases in
which little male DNA is present, as well as in deficient paternity, immigration, and estate cases. Orchid Cellmark has developed a megaplex
for use on the ABI PRISM® 3100 Genetic Analyzer that examines many
Y-STR markers in a single amplification reaction. The Y-STR megaplex
includes all of the European minimal haplotype and U.S. Y-STR haplotype loci, as well as additional, highly discriminating loci. The results
of studies aimed at optimizing the amplification and analysis conditions
for this multiplex will be presented. Validation studies demonstrating
the effectiveness of the Y-STR megaplex as a forensics tool will also be
discussed. In addition, Orchid Cellmark has optimized a Y-STR 10-plex
which has amplicons less than 200 base pairs in length, making it an
ideal system to use with degraded samples containing male DNA. The
present study compares the Y-STR 10-plex and Y-STR megaplex to
determine their ability to effectively discriminate among male lineages.
Y-STR, Forensic, Multiplex
B23
Whole Genome Amplification
of Limited Quantity Samples
Nana Yaa S. Lamouse-Smith, BA, MS*, Robin L. McDowell, MFS*,
Robert E. Wenk, MD, and Francis A. Chiafari, MS*, BRT
Laboratories, Inc., 400 West Franklin Street, Baltimore, MD 21201
After attending this presentation, attendees will learn about a new
technique they may want to incorporate into their SOPs for handling
limited biological samples.
humanity by describing the technique which can be used on challenging
samples to obtain DNA profiles. This may help to obtain DNA profiles
(either complete or partial) from limited samples where before this may
have not been feasible or results achieved did not provide enough information.
This study investigated the utility of Whole Genome Amplification
(WGA) as a method to enhance the sensitivity of current STR technology for samples with limited cell quantity. WGA uses Phi29 DNA
polymerase and random hexamer primers to amplify eucharomatic DNA
by highly processive strand displacement. Our initial experiments
involved determining the minimum number of cells necessary to yield
successful WGA, followed by phenotyping with Profiler Plus and
Cofiler to measure fidelity. Epithelial cells were collected, counted, and
lysed prior to WGA, with the yield determined by Pico Green. Serial
dilutions with as few as 10 total cells yielded WGA product, and produced accurate profiles with Profiler Plus and Cofiler. Initial experiments indicate that this technology may improve the production of profiles from minimal samples in a cost-effective manner. Applications of
this technology to mock and non-probative samples will be discussed.
Whole Genome Amplification, Limited Sample, DNA
34
B24
Guessing the Race From an STR Profile
Charles H. Brenner, PhD*, Consulting in Forensic Mathematics,
6568 Sobrante Road, Oakland, CA 94611
routine DNA profile – the CODIS STR loci for example – properly
evaluated, usually provides strong information about the ancestry of the
contributor.
Calculation of probable racial origin of a crime stain can occasionally be a helpful hint and is extremely easy and cheap to compute.
Therefore, this presentation will impact the forensic community and/or
humanity by providing a tool that the investigator may as well have.
The routine DNA profile – the CODIS STR loci for example –
properly evaluated, usually provides strong information about the
ancestry of the contributor.
It can be useful to know the race or population group of origin of a
crime stain. For example, in April 2003, the Louisiana serial killer
Derrick Todd Lee was arrested based on a tip from a citizen who had,
according to the newspapers, long been inclined to be suspicious of him.
However, the informant decided his hunch was worth acting on only
when and because the authorities, in about March, issued a statement
contradicting the earlier incorrect public assumption that the killer was
Caucasian. Lee is an African-American.
The March announcement (in substance, it was that no race should
be eliminated from consideration) came shortly after a racial analysis by
DNAPrint based on their specially developed set of markers. However,
such a high-tech approach was not essential in this case. In fact I had
been consulted on the case in January and on the basis of the STR locus
profile obtained by the Lafayette and Baton Rouge crime labs provided
essentially the same information. A dragnet, operating on a racial
assumption based perhaps on a profiler’s analysis or perhaps on some
vague witness account, had methodically extracted DNA samples from
800 Caucasians in an attempt to match the crime stain. After calculating
likelihoods for several races – Caucasian, African-American, Japanese,
Korean, Chinese, Hispanic, Vietnamese – I said, “Caucasian is the last
race I’d look at.” (This was a little glib – on the numbers it is really
second last, before Vietnamese.)
The DNA criminalist commonly calculates several “frequencies”
corresponding to the Caucasian, African-American, etc., population
groups. “Frequency” is the wrong word though, since the number is
invariably so small that if it were really interpreted as a frequency it
would imply a fraction of a person. What the number actually represents
is expected frequency, or probability: The probability that a random e.g.,
Caucasian person unrelated to the crime stain would have the crime stain
type. That is, it would if instead of making the customary conservative
computation, we try to be as accurate as possible. To that end, the right
procedure is simply to include (temporarily) the crime stain in the population sample – note that this avoids ever having a probability of zero –
and then use the consequent sample frequency for each allele as a good
estimate of its random match probability. The product rule, perhaps elaborated by the NRC homozygote and heterozygote theta correction (but
with a realistic, i.e., very small, value of theta), is then used to combine
the individual allele probabilities to form a profile probability.
The profile probability assuming Caucasian origin and the profile
probability assuming African-American origin are two probabilities of
the same thing under different hypotheses. As such, their ratio is by definition the likelihood ratio supporting Caucasian over African-American
origin for the source sample. In the Baton-Rouge case, the likelihood
ratio favored African-American over Caucasian by about 6:1. Testing
Caucasians is thus quite inefficient. Curiously, the likelihood for
Japanese, Chinese, and especially Korean populations was even higher,
the likelihood for Koreans exceeding that for Caucasians by a factor of
200. That does not, of course, mean that a Korean was probably the
culprit, only that any single random Korean would be a more plausible
suspect than any single random Caucasian or anything else. Hence
* Presenting Author
lacking some reason to the contrary (such as knowledge that Koreans are
improbable serial killers), the most efficient dragnet would begin with
Koreans. As few Koreans as there are in Baton Rouge – about 0.1% of
the population – the net chance that a Korean would be the killer was
6%. Disproportionately large, but still small. Caucasians and AfricanAmericans, each of which represent about half the people in Baton
Rouge, figured as 11% and 67% respectively to include the killer.
The 6:1 likelihood ratio favoring African-American over Caucasian
seems like a strong clue, but in fact it was a little unlucky it was so small.
On average we can expect to do better. This table shows the typical likelihood ratio that can be expected when comparing various population
groups:
Cau
Caucasian
Hispanic
African American
Japanese
His
5
AA
40
30
Jap
30
10
300
Vietnamese
300
200
5000
8
In the long run forensic STRs will certainly not be the most
accurate tool for assessing racial origin. However, they do offer a
number of advantages. The typing is routine; it doesn’t cost extra.
Extensive population data is available, for a large number of populations, whereas for a specialized test the data will need to be gathered at
considerable expense just for that test. Once it is appreciated that we are
dealing with probabilities and not frequencies, it is apparent that large
population studies are not necessary. Small ones generally are somewhat
less informative, but their smallness does not inhibit inferring a probability. Probability is, after all, a summary of whatever information we
may posess. Finally, the method of analysis of STR data conveniently
gives a likelihood ratio which means that though it may not always give
a definite answer it will rarely be misleading; when it is not sure, it tells
you it is not sure.
Racial Attribution, DNA Typing, Likelihood Ratio
B25
A Review of Mixture Profiles in
the CAP Forensic Identity
Proficiency Testing Program
Leigh B. Thorne, MD*, University of North Carolina Hospitals,
Department of Pathology, 101 Manning Drive, CB# 7525, Chapel Hill,
NC 27514; Robert W. Allen, PhD, 4502 East 41st Street, Tulsa, OK
74137; Arthur J. Eisenberg, PhD, University of North Texas, 3500
Camp Bowie Boulevard, Ft. Worth, TX 76107; David Feldman, MD,
7171 Buffalo Speedway, Houston, TX 77025; Chantal R. Harrison,
MD, 7703 Floyd Curl Drive, San Antonio, TX 78284; Herbert F.
Polesky, MD, 6355 Mountain View Ranch Road, Healdsburg, CA
95448
The goal of this presentation is to review the reported mixture profiles of human spermatozoa with epithelial cells and white blood cells in
proficiency DNA testing results from the Forensic Identity proficiency
testing program.
humanity by demonstrating to the forensic DNA community is doing an
excellent job in the interpretation of mixed samples.
Learning Objective: to review the reported mixture profiles of
human spermatozoa with epithelial cells and white blood cells in proficiency DNA testing results from the Forensic Identity proficiency testing
program.
In 1993, the College of American Pathologists first offered the
Forensic Identity (FID) proficiency testing program. The FID program
distributes two (2) mailings per year. A combination of specimens consisting of reference bloodstains, mock vaginal swabs, crime scene bloodstains and/or semen stains, and other biological material are provided in
each survey. Since a significant percentage of submitted cases to the
* Presenting Author
crime laboratories contain mixed specimens such as vaginal swabs, oral
swabs, anal swabs, or stains from sexual assault cases where human
spermatozoa are present, the College includes such specimens in the
surveys. In every mixed sample to date in the FID program, the contributor of the “cell” fraction has been from a female victim.
One major advantage of DNA testing over traditional serological
testing is the ability to separate the spermatozoa (hereafter referred to as
“sperm”) from the epithelial and white blood cells (hereafter referred to
as “cells”). The first survey of this program contained a mock vaginal
swab sample. The swab was prepared from isolated white blood cells
from the same donor of the victim. The leukocytes were isolated in a
LeucoPrep tube and the leukocytes were counted on a Coulter counter.
Approximately one million white blood cells were spotted onto a sterile
SOLON cotton-tipped applicator and allowed to air dry. Semen from the
suspect was mixed 1:1 semen:1X PBS. 50µL of this mixture was spotted
onto the cotton-tipped applicator. Variations of this original procedure
are still used in this program today. Other mixed specimens have
included a mixture of whole blood and semen on a swab, an undergarment, and filter paper.
In the earlier surveys, participants reported results mostly for
restriction fragment length polymorphism (RFLP) analysis, with some
laboratories reporting results for AmpliType™ HLA DQ-alpha, D1S80,
and a couple short tandem repeat (STR) loci. Today, approximately 150
laboratories are reporting results with STRs in the CAP Forensic Identity
program. The forensic community has standardized the reporting of
PCR STR technology over the years; in-so-much, the survey has evolved
in the reporting of mixed specimens.
Since STR technology is the current standard in forensic laboratories, an evaluation of the separation of these mock mixed specimens
and the reporting of the discrete alleles has been conducted. Not surprising, the “sperm” fraction contained less minor contribution of DNA
from the “cells” due to the ability of the DNA analysts to successfully
separate the spermatozoa from the cell fraction. This presentation will
examine the different combinations of discrete alleles and portray the
combinations of mixtures reported. For example, with >10 participants
reporting, 96.1% to 100% of the participants reported the profile of the
sperm fraction only, representing excellent separation. Of those systems
with 100% of the participants not reporting a mixture, the victim shared
one or more alleles with the suspect and had no alleles different than the
suspect. Further, upon close examination of the “cell” fraction, the different combinations of reported results for this sample demonstrated a
greater range of variation depending on the combination of shared and
non-shared alleles. This combination of alleles will be closely examined.
Pattern interpretation of such mixtures from the most recent surveys
using STR results will be presented.
Proficiency Testing, Mixture Interpretation, College of American
Pathologists
B26
A Safe Methodology to Identify
Biologically or Chemically
Contaminated Evidence
Theresa A. Caragine, PhD* and Linnea Schiffner, MS, Office of the
Chief Medical Examiner, Department of Forensic Biology, 520 First
Avenue, New York, NY 10026; Office of Chief Medical Examiner,
Department of Chemistry, New York, NY 10016; Melissa Liriano,
University of Maryland, Baltimore County, MD 10016; James
Sebestyen, BS, Robert Shaler, PhD, Frank DePaolo, MS, and
Mechthild Prinz, PhD, Office of Chief Medical Examiner, Department
of Chemistry, 520 First Avenue, New York, NY 10016
efficacy of a protocol designed to safely identify, through PCR DNA
testing, the source of blood, semen, and saliva stains, and epithelial cells
recovered from biologically or chemically contaminated evidence.
35
This presentation will demonstrate how irradiation provides a safe
method of decontamination while preserving the integrity of the evidence for PCR DNA testing. Specifically, the methodology limits the
exposure of the evidence collection team to the contaminant and obliterates the risk to laboratory personnel.
In an event that involves weapons of mass destruction, biologically
or chemically contaminated evidence may require PCR DNA analysis
for criminal investigations or body identification. Since most forensic
science laboratories operate at biosafety level two, protocols must be
designed that ensure the safety of forensic examiners, while preserving
the integrity of the evidence. Irradiation at doses adjusted to destroy the
structure and functionality of a pathogen, likely leave identifying DNA
sequences intact. Therefore, evidence samples were irradiated and analyzed by PCR DNA testing, and compared to identical non -irradiated
samples. Most samples, with the exception of low copy number DNA
samples, generated full DNA profiles, although the amount of DNA produced was reduced.
Two identical sample sets were assembled. Samples such as blood,
semen and saliva stains at various concentrations and combinations, cigarette butts, clothing, hair, lip prints on tape, and toothbrush bristles were
equally divided. Alternatively, duplicate samples were obtained from the
same sources for cookie crumbs, cosmetics, ear and eye swabs, razors,
swabs of fingerprints on glass, a metal doorknob, a metal file rack, a
stapler, and a tackle box, and swabs of touched objects such as bottles, cell
phones, credit cards, cups, jewelry, keys, and wallets. Evidence was
packaged within two sealed envelopes or containers and stored within two
boxes that were irradiated intact with 32kGy of energy from an LX 2 High
Power X-ray, courtesy of Titan Inc. Control non-irradiated samples were
similarly stored at room temperature in the laboratory.
Samples from both sets were processed simultaneously. Semen and
mixed epithelial cell samples were extracted by differential lysis using
Chelex beads. According to a modified DNA IQTM (Promega) protocol
for degraded samples, blood and saliva samples were extracted on the
Biomek 2000. Low level DNA samples, for example swabs of fingerprints and touched objects, were digested with 0.01% SDS and
Proteinase K at 56º C for two hours, incubated at 100º C for 8 minutes,
and following the addition of 1mg of Poly A RNA, were purified and
concentrated with a Microcon 100. Samples were quantitated with either
the Quantiblot® kit (Applied Biosystems) or with an ALU based PCR
method with an endpoint determination using a fluorescent plate reader.
1ng or 19.2 µL of each sample were amplified with Promega’s
PowerPlex® 16 kit for 32 or 35 cycles, as needed, and separated on an
ABI 3100 Prism® Genetic Analyzer.
Irradiation reduced the yield of DNA for the majority of the
samples. For example, irradiated blood, semen and saliva stains and
swabs of fingerprints and touched objects contained 68.8%, 64.4%,
97.5%, 54.1%, and 33.4% less detectable DNA than non-irradiated
samples, respectively. Despite this decrease, blood, semen, and saliva
stains were still robust and produced full DNA profiles. However, DNA
profiles for the minor components of blood stain mixtures were compromised. Although 93.8% of the alleles could be deduced correctly for
the minor components in the control samples, only 21.1% of the alleles
were apparent in the irradiated samples. Similarly, 55% of the alleles
were called correctly for control swabs of touched objects whereas only
7.2% of the alleles were designated for the same irradiated swabs.
Samples that have very low copies of DNA, fingerprint swabs, generated
at least 400% less accurate DNA profiles following radiation.
The decrease in DNA yield suggests that irradiation degrades DNA.
Samples with low amounts of DNA are significantly compromised by
radiation, and thus produced partial or no PCR Nuclear DNA profiles.
These low copy number DNA samples would be better candidates for
SNP analysis. However, irradiation does not affect allelic calling when
the DNA recovered is sufficient. Therefore, our study demonstrates that
irradiation may be employed to decontaminant most forensic samples
36
for PCR Nuclear DNA testing. Implementing this described irradiation
process to decontaminate evidence would limit the exposure of the evidence collection team and obliterate any risk to laboratory personnel.
Decontamination, Degradation, Low Copy Number DNA
B27
Separation of Spermatozoa and Epithelial
Cell Mixtures by Laser Microdissection
for Forensic DNA Analysis
Christine T. Sanders, BA* and Daniel A. Peterson, PhD, FUHS/The
Chicago Medical School, Department of Neuroscience, 3333 Green
Bay Road, North Chicago, IL 60064; Nick Sanchez, BS, Los Angeles
Police Department, Scientific Investigation Division, 555 Ramirez
Street, #270, Los Angeles, CA 90012
After attending this presentation, attendees will learn a method for
the selective separation of sperm and epithelial cell mixtures in sexual
assault evidence using Laser Microdissection such that the retrieved
cells can be individually typed for Short Tandem Repeat (STR) analysis.
This presentation will offer the forensic community a new method
for cell separation in challenging sexual assault casework samples.
The goal of this presentation is to present to the forensic community
a method for the selective separation of sperm and epithelial cell mixtures in sexual assault evidence using Laser Microdissection such that
the retrieved cells can be individually typed for Short Tandem Repeat
(STR) analysis.
PCR and STR analysis has become a valuable tool in identifying
the source of biological stains particularly in the investigation of sexual
assault crimes. Difficulties in analysis arise primarily in the interpretation of mixed genotypes when cell separation of multiple donors is
incomplete or when only a small number of target cells are available in
a mixed sample following the application of traditional preferential lysis
procedures.
A typical rape evidence swab may comprise of sperm cells from the
assailant and vaginal epithelial cells from the victim. The differential
extraction has been the most commonly used method to separate sperm
cells from epithelial cells in sexual assault evidence. It can preferentially lyse epithelial cells and/or blood leukocytes cells releasing the
DNA, which can then be removed from the sperm cell pellet. Although
this method can generally provide two cellular fractions, one comprising
of sperm cell DNA and the other of epithelial cell DNA, the separation
is not always complete, and there may be carryover from one fraction to
another making eventual genotype interpretation and further statistical
analysis challenging. This challenge is frequently encountered in cases
where the ratio of vaginal epithelial cells to sperm cells is large or there
are very few numbers of sperm.
Laser Microdissection technology (LMD) has emerged as a method
to capture single cells or a group of cells of interest from heterogeneous
tissue. This technology is typically employed on histological tissue
cryosections to collect specimens for further DNA, RNA or protein
analysis. The purpose of this research is to use LMD on biological
smears to identify both stained and unstained sperm and epithelial cells
while selectively dissecting and recovering the cells of interest for
forensic DNA analysis.
Slide smears comprising of sperm and oral epithelial cell mixtures
were prepared on glass foiled PEN microscope slides. Smears were
processed both stained (Hematoxylin & Eosin or Nuclear Fast Red &
Picroindicocarmine) and unstained.
Automated laser microdissection was performed on the Leica AS
LMD System. The instrument comprises of a Leica upright microscope
with a motorized stage and a single-handed xyz control. Coupled with
a camera and computer software, target cells can be visually identified
and selected on the computer screen for microdissection. The microscope is integrated with a UV laser of 337µ which performs the laser
* Presenting Author
ablation of the plastic PEN film around the cell(s) of interest. The cut
material is then deposited automatically into one of the designated PCR
tubes. An inspection mode allows the confirmation and image documentation of the targeted cells in the PCR tube before further molecular
analysis.
The Laser Microdissection method presented physically dissects
target cells without the contamination of adjacent foreign cells in a
mixture then collects the target cells for direct DNA isolation. This
bypasses the multi-step process of a preferential lysis procedure, which
can reduce yield through loss by liquid transfers and premature lysis of
target cells. Unsuccessful analysis of sexual assault evidence due to low
numbers of sperm cells or a large epithelial/sperm cell ratio could
instead be subjected to LMD to provide effective cell separation and
maximal cell yield.
Laser Microdissection, Forensic DNA Analysis, Sexual Assault
Evidence
B28
Development of a Microfabricated Device
for Separation of Sperm and Vaginal
Epithelial Cells: A Significant Step
Toward Circumventing Conventional
Differential Extraction
Katie M. Horsman, MS*, Katherine A. Koen, and Jerome P.
Ferrance, PhD, University of Virginia, Department of Chemistry,
PO Box 400319, Charlottesville, VA 22904; James P. Landers, PhD,
University of Virginia, Departments of Chemistry and Pathology,
PO Box 400319, Charlottesville, VA 22904
The goal of this research presentation is to introduce microchip technology to the forensic community and demonstrate its potential for the
separation of sperm and vaginal epithelial cells in rape kit analysis.
This presentation will introduce the forensic community to microchip
technology and its potential for rapid analysis of rape kit DNA evidence,
thereby, reducing the DNA analysis backlog.
Differential extraction, the conventional method for isolating male
and female fractions of DNA, is a time-consuming sample preparation
step in the forensic DNA analysis of rape kit evidence. Therefore, our goal
is to develop a means to reduce the time associated with isolation of the
male and female DNA fractions, while maintaining or improving the
percent recovery and purity. The means through which we are attempting
to achieve this goal is the use of microfabricated glass devices.
The brief record that exists for microminiaturization of analytical
processes on microchip platforms has demonstrated that reduction in
analysis time (versus conventional methods) is often a benefit, as well as
the potential for integration of multiple processing steps in a single device
and for automation of these processes. Since differential extraction is only
one of a number of processes that constitute forensic DNA analysis,
replacing it with a microdevice method provides a distinct advantage with
the possible integration of several sample preparation steps, including
DNA extraction, DNA quantitation, and PCR amplification on a single
device. In addition, microchips can be designed to accommodate parallel
processing of both the male and female DNA fractions as well as the necessary positive and negative controls. While an integrated microdevice
including all sample preparation steps is advantageous in many respects,
the cell separation methodology is amenable to a modular system in which
separate devices are developed for each processing step, depending upon
the needs of the forensic community.
The conventional differential extraction methodology is not easily
translated to the microchip format because of the centrifugation and filtration steps. Therefore, a novel method for obtaining isolated male and
female fractions of DNA on a microfabricated device was developed. This
new technique involves first separating the sperm cells from the cell
mixture, then extracting the DNA from each fraction independently,
allowing separate male and female DNA fractions to be obtained.
* Presenting Author
The cell separation step is the focus of the research presented here.
The separation developed exploits the differential physical properties of
the two cell types such as buoyant density, size, shape, and proclivity for
adhesion to the microchannel surface. In an etched microchannel, a flow
rate of approximately 1 nL/sec was obtained using a mechanical pump,
directing the sperm cells to a collection reservoir while retaining the
epithelial cells in the inlet reservoir. Preliminary experiments employed
digital video microscopy to visualize the cell separation and demonstrate
the purity and efficiency of the process. Methods have been established to
selectively separate free DNA (from the more easily lysed epithelial cells)
from the sperm cell fraction. The movement of bacteria during the cell
separation has been characterized, although the presence of bacteria is not
particularly concerning because of the lack of amplifiable human STRs.
In addition, the movement of white and red blood cells must be characterized, because of DNA contamination and PCR inhibition issues, respectively.
Using mock post-rape vaginal swabs, the cell separation product
obtained on the microdevice resulted in a clean sperm cell fraction. The
DNA from isolated cells was extracted with a commercial extraction kit,
amplified with a Profiler® PCR kit, and analyzed on an ABI 310 commercial CE, yielding the profile of the male sperm donor. DNA extraction
from the isolated cells was also performed using a microdevice method
before amplification and analysis to demonstrate the potential of integrating these two steps into a single device to circumvent conventional differential extraction.
Differential Extraction, Cell Separation, Microchip Technology
B29
Decontamination of Human Autopsy
Specimens by 60Co Gamma-Photon
Irradiation and Human DNA
Identification by Short Tandem
Repeat Analysis of Irradiated Tissues
Susan W. Jones, PhD, MFS* and Demris A. Lee, MSFS, Armed
Forces DNA Identification Laboratory, 1413 Research Boulevard,
Building 101, Rockville, MD 20850; Thomas B. Elliott, PhD,
Michael O. Shoemaker, PhD, and Gregory B. Knudson, PhD, Armed
Forces Radiobiology Research Institute, 8901 Wisconsin Avenue,
Bethesda, MD 20889-5603; Brion C. Smith, DDS, Armed Forces DNA
Identification Laboratory, 1413 Research Boulevard, Building 101,
Rockville, MD 20850; Craig T. Mallak, MD, Armed Forces Medical
Examiner System, 1413 Research Boulevard, Building 102, Rockville,
MD 20850
After attending this presentation, the participant will understand 1)
the use of Cobalt 60 (60Co) gamma-photon ionizing radiation for decontaminating human autopsy specimens and 2) the effect of decontaminating
doses of gamma radiation on Short Tandem Repeat (STR) DNA analysis
of tissue and bone specimens.
This presentation will impact the forensic community by demonstrating irradiation as a way to decontaminate human remains found in an
incident involving contaminating weaponized microbial agents prior to
submission the laboratory for human identification by STR DNA analysis.
Short Tandem Repeat (STR) DNA typing has been the primary tool
for identification of human remains in military and mass disaster incidents. The majority of the military specimens submitted to the Armed
Forces DNA Identification Laboratory (AFDIL) include fresh tissues and
bones collected by medical examiners during autopsies.
The potential use of military force in nations that may have biological
weapons of mass destruction, such as weaponized Bacillus anthracis
spores, and the potential for large numbers of fatalities from inhalation
anthrax in these nations suggested that we study a method for decontaminating human remains. We investigated 60Co gamma-photon irradiation as
a method of decontamination.
37
In this study, 11 different human cadaveric finger specimens were
collected from 6 female and 5 male deceased donors. DNA was organically extracted and then quantitated using the Quantiblot® method. STR
profiles were obtained from a 3- to 5-mm piece of tissue from each of the
donor fingers for a known DNA STR typing control prior to gamma irradiation. The organic DNA extraction and Profiler Plus™ STR profiling
methodologies are rapid, reliable and robust technologies that have been
validated for casework at AFDIL, accepted in the forensic community, and
published in the forensic literature.
A STP-350 biological shipment container (SAF-T-PAK, Edmonton,
AB, Canada) was chosen to safely contain biological specimens, endure
the rigors of transport from a military theater of operation and the process
of irradiation, and meet the biohazardous materials transportation regulations. The large orange suitcase-like shipment container has the internal
capacity to fit eight biohazard canisters in a fitted polystyrene holder. The
biohazard canisters can hold up to five specimen transport tubes and
absorbent paper.
Each of eight specimen transport tubes contained 0.5 g of dry, freeflowing Bacillus subtilis var. niger (B. globigii) spores, i.e., biological
indicators (BI), as a surrogate for B. anthracis, at a concentration of 5.5 ×
1011 colony-forming units (CFU) per gram. A radiation dosimeter (Dalanine pellets) was placed in three of these tubes with the spores. Each of
the 11 donor fingers was placed in a specimen tube containing 99.99% isopropanol. The BI’s and the tissue specimens were then distributed in the
eight canisters in mapped locations within the case.
Gamma irradiation of the shipment container and dosimetry were
performed in the Cobalt-60 Irradiation Facility at the Armed Forces
Radiobiology Research Institute. Electron paramagnetic resonance (EPR)
dosimetry measurements of irradiated alanine pellets were performed at
the National Institute of Standards and Technology (NIST, Gaithersburg,
MD) using published procedures to determine the dose delivered. The
case and contents were given three fractionated doses of gamma-photon
radiation at a constant rate in order to achieve a decontamination assurance
level (DAL) for the BI. The DAL is the dose of gamma radiation required
to kill all spores in 99% of the BIs. Before and after irradiation, BI spore
viability was determined by standard microbiological methods as defined
in the 2002 United States Pharmacoepia 25th edition (USP25).
The gamma-photon radiation dose, which was delivered inside of the
case, was an average of 51.7 kGy as measured by alanine dosimetry and
was sufficient to kill all spores in test BI samples. DNA isolated from 2gram bone specimens and 3- to 5-mm cubed tissue specimens from each
of the irradiated fingers yielded full Profiler Plus™ STR profiles that were
consistent with the STR profiles obtained from the original non-irradiated
donor source fingers. Mutations, extraneous alleles, and allelic dropouts
were not observed in the Profiler Plus™ STR DNA profiles from the irradiated finger tissues and bones.
Conclusions: This study shows that gamma-photon irradiation
decontaminates potentially contaminated human specimens and that the
process does not adversely affect STR DNA analyses for human identification purposes.
Short Tandem Repeat DNA, Human Identification, Radiation
B30
Real-Time PCR Methods for
Analysis of Forensic Samples
Eric Buel, PhD* and Janice A. Nicklas, PhD, Vermont Forensic
Laboratory, 103 South Main Street, Waterbury, VT 05671
After attending this presentation, attendees will understand rapid
quantitative and inexpensive tests for determination of sample DNA
quantitation and gender determination
This presentation will demonstrate that new methods will be a
valuable addition to DNA analysis by saving time and money and
improving the quality of human DNA quantitation.
38
DNA isolated from crime scene samples must be quantitated to
determine the amount of human DNA present. Recently the forensic
community has become interested in developing faster, cheaper and
more quantitative methods than the widely used slot blot method utilizing a D17Z1 probe. In most molecular diagnostic laboratories, PCR
techniques have almost completely replaced Southern and slot blot
methods. Thus, development of quantitative PCR methods to measure
the amount of human DNA in the forensic setting is a logical extension
of this progression. Obviously the sequence chosen must be human or
at least primate specific and if present in multiple copies would allow
more sensitive quantitation. Alu sequences are primate-specific and are
found in ~800,000 copies in the human genome and make an excellent
marker for human DNA.
Our initial assay used endpoint PCR quantitation with SYBR Green
I dye in a fluorescence plate reader. We then turned to a real-time PCR
assay using SYBR Green I. This assay has the advantage of a wider
range, 16ng down to 1pg, a turnaround time of ~87min (with less handson time than the plate reader assay) and a cost of $0.50 per sample. Both
of these methods were optimized by adjusting primer concentration,
annealing and extension time and temperature, SYBR Green concentration and polymerase concentration. The assay was validated by comparison with usual slot blot method on mock and real crime samples and
by STR analysis based on assay quantitation results. The studies
indicate that the SYBR Green Alu method is at least as cost effective and
sensitive as the slot blot method, as well as much simpler to perform and
with a greater analytical range.
PCR quantitation methods that use probes (oligonucleotides with
fluorescent dyes attached) have the advantage that plateau effects are
lessened and a greater dynamic range can be achieved. We have also
investigated and compared the use of LUX primers (Invitrogen), MGBEclipse (Epoch Biosciences) and TaqMan (ABI) probes for readout.
The LUX system has a low fluorescence output and appears not to be
sufficiently reproducible. The Eclipse system has been optimized and
validated with case work samples and can detect from 256ng down to
1pg of DNA at about the same cost as the SYBR™ Green assay. The
TaqMan assay can also detect from 256ng down to 1pg DNA; optimizations and validations are underway.
In addition to simply quantitating the total human DNA in a sample,
it is often of importance to determine whether and how much male DNA
the sample contains. In crimes involving violence against women perpetrated by men, blood may be shed by the both the victim and attacker;
a method that could quickly identify which stains are male or female
could allow the examiner the chance to be selective in the stains that are
analyzed further by STR analysis. Another important use of sex typing
would be in the STR analysis of sexual assault cases where the percentage of male DNA in differential extractions can vary widely. A
method that could determine the amount of male DNA present in a
sample could allow the forensic scientist to decide if a sample has ample
male DNA for autosomal or Y STRs, respectively. We have tested LUX
primers, MGB-Eclipse and TaqMan based Y satellite (Y specific) assays.
These assays are able to discriminate male and female DNA over
100,000 fold and accurately quantitate male DNA. Experiments are
underway to optimize these assays and pick the best assay from among
these methods. The plan is then to develop a multiplex system of gender
determination in conjunction with human Alu DNA quantitation. Such
an assay will greatly reduce the work and expense in analysis of crime
scene samples by quickly determining which samples are important and
which contain sufficient DNA of the correct gender to yield valuable and
conclusive STR results.
Supported under Award number 2000-IJ-CX-K012 from the Office
of Justice Programs, National Institute of Justice, Department of Justice.
Points of view in this document are those of the authors and do not necessarily represent the official position of the U.S. Department of Justice.
Human DNA, DNA Quantitation, Real-Time PCR
* Presenting Author
B31
Extraction and Quantification of Human
DNA and the Amplification of Human
STRs and Amelogenin From Fly Larvae
Found on Decomposing Tissue
George J. Schiro, Jr., MS*, Acadiana Criminalistics Laboratory, 5004
West Admiral Doyle Drive, New Iberia, LA 70560
After attending this presentation, attendees will learn that fly larvae
found on decomposing human tissue could be an alternative means of
identifying human remains.
This presentation will provide support of the forensic science community and the popular media hypothesis that human nuclear DNA can
be recovered from fly larvae found on human remains.
The purpose of this experiment was to determine if human deoxyribonucleic acid (DNA) could be extracted, quantified, and amplified
from fly larvae that had fed on decomposing human tissue.
Blowfly (Diptera: Calliphoidae) eggs were collected and placed on
donated human skin, muscle, or liver tissue in rearing chambers. After
two to twelve days, the larvae were collected in 70% ethanol, 2propanol, acetone, or by freezing. One to sixteen larvae were washed
and dissected by mincing, splitting, or bisecting the larvae. The DNA
was extracted using digest buffer and either phenol: chloroform: isoamyl alcohol (PCIA)or by using silica spin columns. The human DNA
was quantified using a biotinylated primate specific probe. The DNA
extract was then amplified using the polymerase chain reaction (PCR)
for 28 cycles and primers for 13 specific Short Tandem Repeats (STRs)
and the sex identification marker amelogenin. An additional seven PCR
cycles were used on extracts that failed to produce any amplified human
DNA. Washes from the exterior of the larvae that produced amplifiable
human DNA were also analyzed. Finally, the PCR product was separated
and characterized using capillary electrophoresis.
Out of the 30 DNA extracts, one sample had approximately 4.59.0ng of human DNA. The remaining 29 extracts were either void of
human DNA or the amount of human DNA fell below the test detection
limits. Three of the thirty extracts produced amplified products for the
sex identification marker and seven to eight STRs. The STR profiles
produced by these three extracts matched the STR profiles of the corresponding positive controls. Some of the extracts subjected to the seven
additional PCR cycles provided limited and often inaccurate identifying
information. The experiment also provided evidence, but not conclusive
proof, that the human DNA originated from inside the larvae.
The following conclusions were drawn from the experiment:
Human tetranucleotide STR analysis is highly sensitive and selective.
Macromolecules survive larval ingestion. Three to four larvae are
required for this type of analysis. Muscle and liver tissue are the best
feed tissues from which to collect the larvae. Digest buffer and PCIA are
more suitable than silica spin columns for recovering human DNA from
fly larvae. 2-propanol is a better collection medium than 70% ethanol,
acetone, or by freezing for this type of analysis. Mincing or splitting the
larvae provided better results than bisecting the larvae and using the
anterior portion of the larvae. The drawbacks of additional PCR cycles
outweighed any additional benefits.
Larvae, DNA, STRs
B32
The Comparison of Drug Chemistry
Training Programs of the Illinois State
Police, University of Illinois at Chicago
Residency and the National Forensic
Science and Technology Center
Richard A. Paulas, BS*, Illinois State Police, 1941 West Roosevelt
Road, Chicago, IL 60608
This paper describes the programs and goals of each program and
provides some measurable data for comparison of their effectiveness.
This presentation will provide a description of various chemistry
training programs and their financial impact upon the host.
The Illinois State Police Forensic Sciences Command has conducted training for its forensic scientists for more than 25 years and has
earned an excellent reputation. Their program is recognized nationally as
providing both the theoretical and practical knowledge necessary to
conduct quality forensic analysis before they are required to work actual
cases. The Chemistry Training program recently had the opportunity to
compare the results of training from two other training programs with its
own. In January of 2002, six individuals were hired by the Illinois State
Police, Forensic Sciences Command to be trained in Drug Chemistry
analysis. Two individuals were asked and agreed to be trained by the
National Forensic Science and Technology Center (NFSTC) in Largo.
Florida. Three individuals proceeded in the normal drug chemistry
training program at the Forensic Science Center in Chicago. The final
individual was completing his residency requirement as part of his
Masters of Forensic Science from the University of Illinois at Chicago.
This paper describes the programs and goals of each program and provides some measurable data for comparison of their effectiveness.
Chemistry, Training, Residency
B33
Institutional Support for Forensic Science
Educational Programs: A Survey
Jay A. Siegel, PhD*, Michigan State University, School of Criminal
Justice, 506 Baker Hall, East Lansing, MI 48824
After attending this presentation, attendees will learn how forensic
science educational programs are supported by their institutions and
learn ways to help improve the funding situation
This presentation will offer to the forensic science community, for
the first time, information about how our forensic science education programs are faring financially and where the gaps are and what can be
done to help in the future.
There has been a lot of discussion at AAFS meetings and at other
forums about how poorly forensic science education programs are
funded relative to other science based programs at US colleges and universities. There is not enough money for faculty, instrumentation,
graduate assistants, grants, research, etc. But what is the real situation?
How are forensic science educational programs funded and how well? A
survey was conducted among 46 colleges and universities that self identified as having a bachelors degree or masters degree program. The
survey looked at staffing levels, graduate student support, equipment,
travel and grant and research support. The results are presented along
with implications and suggestions for the future.
Education, Research, Funding
* Presenting Author
39
B34
Premarital Sex, Schizophrenia, and
Other Rape Victim Hazards
Karolyn L. Tontarski, MS*, Montgomery County Crime Laboratory,
2350 Research Boulevard, Rockville, MD 20853
After attending this presentation, attendees will recognize scenarios
that may be preventing the submission of physical evidence in rape and
assault cases
This presentation will impact the forensic community by demonstrating how to recognize investigative practices that may be limiting
effective sexual assault investigations, the value of utilizing forensic
biology evidence to identify cases needing further investigation, and
educating forensic biologists on the important role they can play in these
potentially compromised investigations
Given the nature of the crime, rape is often seen as difficult to
investigate and prosecute. Typically, rape victims fall into one of two
categories – stranger assault victims or acquaintance assault victims.
Society seems to sympathize with victims of stranger rape more. Indeed,
once the perpetrator has been identified there is typically little, if any,
reluctance to prosecute these cases. Acquaintance assault cases can be a
bit more problematic – as can stranger assault cases with “compromised” victims. Victims who have affected their personal safety often
put off investigators, prosecutors, and jurors, as do victims who are perceived to be problematic. It is not uncommon to hear that a case won’t
be prosecuted because the victim is a prostitute, mentally ill, used drugs,
etc. Nor is it uncommon to hear that a defendant was found not guilty
because a victim “left the bar willingly with him” or “should not have
been drinking so much.”
It is specifically because of these sorts of issues that all forensic
biologists need to understand, at the very least, the dynamics of rape.
Without foundational knowledge regarding the nature of sex crimes, the
somewhat negative environment that is associated with rape can
adversely affect forensic biologists. If the forensic scientist is to remain
impartial, he or she must be able to understand the context within which
these comments are made. While it should be possible for forensic biologists to be insulated from some of these issues, the reality is that sexual
assault investigators and attorneys often look to forensic biologists for
guidance with regard to what evidence should or should not be
examined. Examiners with a lack of understanding as to what rape is
can potentially be confused as to what evidence needs to be processed.
In the worst case scenario, they may advise that evidence they perceive
to be non-probative should not be processed.
Given the premise that some women are more easily victimized
(such as prostitutes, mentally ill persons, etc.), it stands to reason that
certain types of rapists will hone in on them. Typically, these women are
less likely to report the crime, or if they report it, they are less likely to
be believed. When these victims are discounted by the system, we are
making it easier for these sorts of crime to continue. It is not only possible, but is highly probable, that these women are included amongst
many serial rapists’ victims. This issue has become relevant to the
unsolved case DNA backlog reduction project.
With the advent of DNA testing, there has been a push to test backlogged rape evidence. In cases where this evidence has been examined
and data bank hits have been obtained, there can be a failure of cases
with “compromised” victims to move forward in the criminal justice
system. In some cases, decisions are being made to not even conduct
testing on cases with “compromised” victims. This presentation will
address the need for educated forensic biologists to actively participate
in defining what cases should be processed. In addition, some discussion will center on working with investigators and attorneys to
encourage them to fully consider these difficult cases, and the value of
prosecuting these crimes.
Rape, Sexual Assault Investigation, Victims
40
B35
Unique Solutions to Addressing the
Backlog of Criminal Casework
Michael L. Cariola, MFS*, Todd Bille, MS, James Schumm, PhD, and
Mitchell Holland, PhD, The Bode Technology Group, 7364 Steel Mill
Drive, Springfield, VA 22150
By attending this presentation, attendees will learn of solutions
developed to effectively address the backlog of cases requiring DNA
analysis.
humanity by demonstrating the combination of a growing backlog of
cases requiring DNA analysis and the severe budget constraints that
states are facing requires novel approaches to the analysis of casework.
The authors want to share with the community the processes developed
to help address this problem.
Crime labs across the country face a growing backlog of cases
requiring DNA analysis. This includes not only an increased demand for
DNA-testing on current cases but also the testing of older cases which
were either never submitted due to the lack of a suspect or were tested
prior to the adoption of the 13 CODIS STR loci. This has left many
crime labs with hundreds of ‘no-suspect’ cases requiring analysis but no
increase in the number of analysts available to do the work. New York
City alone had a backlog of greater than 15,000 no-suspect sexual
assault kits. From October 2000 to July 2003, we analyzed over 6000 of
these kits for the New York City Police Department. During this time,
we initiated work with many other crime labs with different requirements requiring customized solutions to their problems. In order to
process these cases in the most efficient manner and meet the demands
of various users, we have developed processes addressing the primary
bottlenecks encountered during the analysis of these cases. The primary
areas covered here include 1) the screening of the kit to identify male
component DNA, 2) quantitation of DNA extracts and 3) final generation of a CMF file and a court-ready allele table.
One of the primary bottlenecks to the analysis of sexual assault
cases is the initial screening of a case for the presence of seminal fluid.
Typically, this entails a combination of Acid Phosphatase (AP)
screening, microscopic searches for spermatozoa and/or P30 testing. In
a typical sexual assault evidence collection kit containing multiple
vaginal, anal, oral and various body swabs, it can take over an hour to
screen a single kit. Rather than use a traditional serology approach to
screening, we have developed a DNA-based procedure in which we differentially extract one sample of each type without any pre-screening.
Following DNA extraction of case samples, we employ a Y-Marker
Screen (YMS) to screen samples for the presence of male DNA. The
YMS test gives a greater level of profile sensitivity than the STR multiplexes used for DNA analysis, so false negative results are eliminated.
The YMS test consists of the Amelogenin primers as a control to ensure
that either X-chromosome or both X and Y-chromosome DNA are identified (if only X-chromosome DNA is observed, then only female DNA
is present). Primers for two monomorphic Y-Sequence Tagged Sites
(STS) are co-amplified with the Amelogenin primers. The STS’s are
only diagnostic for the presence of Y-DNA, and do not require STR-like
analysis. As a result, quantification is not required prior to YMS-amplification. Additionally, the amplitude of the STS product is diagnostic for
the approximate quantity of male DNA in the sample extract, providing
value to the scientist when attempting to troubleshoot complex STR
analysis results. Given these characteristics, the YMS test provides a
simple and efficient way to determine if male DNA is present and if an
STR profile can be generated from a sample. Samples that test negative
for the presence of male DNA at this point can be reported as negative
and not processed further. This system has been used on over 3500 cases
and our data suggests that it has resulted in a higher percentage of
foreign profiles for eventual CODIS upload than traditional screening
approaches.
* Presenting Author
Once positives have been determined, all samples must be quantified using a primate-specific assay. Rather than using a traditional
approach such as a hybridization assay, we have developed a PCR-based
assay which amplifies a defined region of the human TH01 locus with
subsequent detection using PicoGreen on a fluourometer. This gives a
precise representation of human DNA present in the sample in an output
format (Microsoft Excel) that allows for rapid calculation of dilutions
before amplification. Equally important, because the system uses simple
commercially-available primers and a low-cost detection system, the
cost for quantitation is much lower than other commercially available
methods. To date, the method has been used successfully on more than
30,000 samples.
The ultimate goal of no-suspect cases differs from traditional
casework. Rather than the generation of a profile for comparison to
known individuals, profiles are generated primarily for upload to
CODIS. While Genotyper is well-equipped for reference samples to
export data in a CMF-ready format, evidentiary samples often contain
mixtures that must be interpreted or partial profiles that must be analyzed locus by locus and across numerous amplifications. Following
analysis of STR data by two independent reads and final review, the data
is traditionally summarized by the hand-entering of an allele table in a
report or the hand-entry of a profile for upload to CODIS. This allows
for potential transcriptional errors and, in the process, creates a
tremendous review burden for the crime lab. To address this problem,
we have written a program for generating CMF files and allele tables
which converts Genotyper table data into both a court-ready allele table
(in Microsoft Word format) and a CMF file (in Microsoft Excel format)
for upload to CODIS. It is customized to the individual laboratory to
meet their review and upload requirements.
The backlog of no-suspect cases across the country is currently estimated at over 300,000 and growing. As states face increasing budget
constraints, the time required to train more analysts will delay our ability
to solve these crimes. By addressing existing bottlenecks in the processing of cases with effective high-throughput solutions, we have successfully increased the overall efficiency of the laboratory while also
increasing the quality of the work. The processes outlined above will be
discussed.
DNA Backlog, No-Suspect Casework, CODIS
B36
2002 Forensic Crime Laboratory
Survey Results
Joseph L. Peterson, DCrim*, Criminal Justice Department, University
of Illinois at Chicago, 1007 West Harrison Street, Chicago, IL 60607;
Susan H. Johns, MA, Illinois State Police, Division of Forensic
Services, 630 East Washington Street, Springfield, IL 62701-1304;
Matthew Hickman, Bureau of Justice Statistics, 810 Seventh Street,
NW, Washington, DC 20531; Tiffany Vasquez, University of Illinois at
Chicago - Forensic Science Program, 210 West Eugenie Street #1N,
Chicago, IL 60614
After attending this presentation, attendees will have received an
updated overview of management, caseload, and resource needs of the
nation’s crime laboratories.
This presentation will provide laboratories and policy makers with
current management and resource needs information about the nation’s
publicly funded crime laboratories.
The Center for Research in Law and Justice and the Survey
Research Laboratory at the University of Illinois at Chicago (UIC), in
concert with the American Society of Crime Laboratory Directors
(ASCLD), were funded by the Bureau of Justice Statistics (BJS) to
undertake a census of all publicly funded crime laboratories throughout
the United States. In addition to gathering baseline statistical information on operations and workload, BJS also wanted us to assess where
added resources were needed. In addition, unlike earlier surveys, the
* Presenting Author
goal was to reach 100% of the nation’s publicly funded crime laboratories. UIC and ASCLD received funding to undertake this research,
given their experience in conducting surveys and studies of crime laboratories over the past twenty-five years.
Staff worked extensively with ASCLD and the community of crime
laboratories to develop a comprehensive listing of laboratories. A draft
version of the survey was designed and pre-tested in selected jurisdictions. All names and addresses were verified via telephone contacts.
The survey itself had six primary areas: Organization, Budget, Staff,
Workload, Outsourcing, and Quality, Training, & Research. The first
section was designed to determine the political entity and population of
jurisdictions served, and types of forensic functions performed in each
laboratory. Next, we examined the budget breakdown of each laboratory, and sources of funding. To gain a greater sense of the crime laboratory workloads, we queried labs about the number of cases received
and backlogged, and the volume of requests received, worked, and backlogged in 2002. We also asked about the performance expectations that
laboratories had of various examiner specialists, and what added
resources would be needed to reduce backlogs. Questions were asked
about the number of requests outsourced (e.g., controlled substances,
toxicology, DNA/CODIS), as well as the cost and funding sources for
such testing. Lastly, we asked if laboratories were accredited, the types
of proficiency tests in which they were engaged, and the dedication of
resources to training and research.
UIC will provide BJS with a documented, formatted, and
archivable data set, and BJS will publish and disseminate a Bulletin
describing results. Project staff, as well as BJS and ASCLD representatives, will be available to answer questions.
Crime Laboratories, Survey, Management and Resources
B37
Where Are the Scientists?
John J. Lentini, BA*, Applied Technical Services, Inc., 1190 Atlanta
Industrial Drive, Marietta, GA 30066
After attending this presentation, attendees will understand the need
for scientists on the crime or fire scene. This will be one of five presentations in a two-hour discussion on this topic.
This presentation will stimulate some thought about how to solve
the problem of fire investigations being conducted by individuals with
little or no scientific training, and to stimulate some interest on the part
of forensic scientists in becoming involved in fire investigations.
Fire investigation is a forensic science that is a world unto itself.
The investigator who ventures here risks exposure not only to a unique
and possibly dangerous physical environment, but to scientific, professional and personal challenges not found in any other field of forensic
science. Scientists from traditional forensic science laboratories may
feel overwhelmed and unprepared for the analytical challenges, many of
which make mastery of a laboratory analysis seem like child’s play. At
first blush, it seems like nothing we learned in our study of chemistry or
physics will explain the chaos that presents itself after nearly every fire,
but with patience, and practice, and a careful application of the scientific
method, the truth can usually be teased out of the ashes.
Unlike a homicide or a robbery or almost any other incident that
requires investigation, a fire is unique in that the first major task, and
often the most daunting one, is to determine whether a crime has taken
place. It is necessary to look pretty carefully to find another field of
investigation in which this is true. Unexplained death comes to mind,
but in that case, there is a clearly defined set of protocols, the forensic
autopsy, that will usually resolve the question unequivocally. The
medical analogy is a useful one because the fire investigator is called
upon to perform a forensic autopsy of a structure or vehicle to determine
the cause of the fire. The deviations from the analogy, however, are what
make it interesting.
41
The medical examiner performing the autopsy has an undergraduate degree, usually in a natural science, a four-year medical education, and several more years of internship and residency in pathology
and/or forensic medicine. The fire investigator, on the other hand, may
have no education beyond high school, and a forty-hour “basic arson”
school, followed by an eighty-hour “advanced arson” school, and continuing education taught by people with the same training, and more
experience. Certainly there are many skilled fire investigators who can
and do perform careful, science-based investigations, even without the
benefit of formal scientific training, but that is certainly not the rule.
The methodology of the medical examiner is also likely to be very
predictable, in that he or she will follow a written, peer-reviewed protocol, and will feel no discomfort at publishing and sharing the results of
the autopsy with colleagues. The methodology of the fire investigator,
on the other hand, depends almost entirely on who the investigator is and
by whom he or she is employed. There is a constant, and frequently
fierce debate on what standards, if any, fire investigators should be held
to, on whether fire investigation is an art or a science or a mixture of
both, and on the level of training and certification required to do this difficult job.
How did this situation come about? How is it possible that an individual with no formal scientific training in chemistry and physics, and no
certification, filled with misconceptions about the phenomenon he professes to have expertise in, is allowed to opine before a jury on issues of
life and death? More importantly, what can be done about it?
This situation arose, quite simply, by default. Comfortable in clean,
air-conditioned laboratories, only occasionally venturing out into a crime
scene, forensic scientists, with few exceptions, have left the field of fire
scene investigation to the non-scientists. They have been content to participate in the modest task of determining whether a sample of debris contains ignitable liquid residue. While proper chemical analysis is
important, and improper analyses have “verified” otherwise unsupportable
hypotheses, the bulk of the hypothesis formation and testing (when the
investigator chooses to follow the scientific method) takes place in the
field, in the dark, dirty, smelly burned out hulks of former residences,
offices and factories. One of the purposes of this session is to encourage
interested scientists to overcome their aversion to disorder and bring their
scientific talents and knowledge to a field sorely in need of it.
But stepping into the field without proper training in the investigation of fire may have worse results than going forth without the benefit
of a scientific education. The current occupants of the fire investigation
community do not yield their territory gladly, and a scientist not affiliated with a federal agency may find actual hostility from local law
enforcement investigators. If scientists are to take over the lead in the
determination of fire causation, they need to be at least as thoroughly
trained as those whom they would replace. They need to go to “arson
school,” to light fires and watch them burn, and to conduct experiments.
The forensic science community needs to find the will to do this job that
has so far been left to non-scientists. It needs to find the money to
provide the necessary training. It needs to find the scientists willing to
get their hands dirty, and it needs to introduce those scientists to the field
in such a way that they are welcomed as legitimate and necessary
participants in the investigation.
Fire Investigation, Scientific Method, Training
B38
Forensic Scientists at the Crime
Scenes—Past, Present, and Future
Richard E. Tontarski, BA, MFS*, Bureau of Alcohol, Tobacco,
Firearms and Explosives, Fire Research Laboratory, 6000 Ammendale
Road, Ammendale, MD 20705
After attending this presentation, attendees will understand that
what may be eroding the role forensic scientists could be playing in the
criminal justice system.
42
This presentation will demonstrate how to begin thinking about the
implications to crime scene processing and forensic scientists’ criminal
justice role
The Bureau of Alcohol, Tobacco, Firearms and Explosives has utilized scientists at major fire and explosion scenes since the late-1970s.
They are an integral part of ATF’s National Response Teams (NRT) and
have played important roles in major investigations ranging from the
recent DC sniper investigation to overseas terrorist bombings.
At these major scenes the scientist, often a chemist, plays many
roles. He/she assists in the scene processing, provides technical information about materials at the scene and most importantly is an advisor
to the team supervisor and a consultant to the team members. Examples
of these roles will be discussed.
ATF’s NRTs are continually evolving to incorporate new technologies that include crime scene mapping, and other technical and scientific specialties. One of the newest additions to the NRTs has been
engineers with a variety of specialties…fire protection, mechanical, and
electrical. The Fire Research Laboratory’s mission is to support fire
investigations in the field, as well as the laboratory. The FRL scientists
approaches will be discussed and illustrated.
What about the other 99% of the cases? What is the role of the laboratory scientist versus that of the crime scene technician? One does not
replace the other. What is the appropriate quality assurance standard
required for scientists working in the field? To which field role do the
standards apply?
Are we inadvertently restricting the role that forensic scientists
play? Misinterpretation or misunderstanding of accreditation and certification standards may be restricting the effective use of forensic scientists. Laboratory management may be unintentionally eroding scientists’
roles, both in the laboratory and in the field, and reducing highly skilled
examiners to technicians. Once there is a perceived limitation about
what role the laboratory examiner should be playing an insidious erosion
in what the scientist is able or qualified to perform begins.
How do we maximize the role of the forensic scientist to provide
bulletproof evidence in court, as well as provide forensic intelligence to
support investigations in a way that only a scientist can?
Crime Scene, Forensic Scientist, Laboratory
B39
Private Consultants for the Prosecution—
Are They Really Necessary?
Peter R. De Forest, DCrim*, Professor of Criminalistics, John Jay
College of Criminal Justice, 445 West 59th Street, New York, NY 10019
After attending this presentation, attendees will have increased
awareness concerning the essential role that comprehensive physical
evidence interpretations play in a complete forensic science service.
The author has served as a private consultant for the prosecution
(and defense) for more than three decades. On a significant number of
occasions this was critical for, or contributed to, the case solution. The
question arises, why was this involvement of a private expert necessary?
The primary answer is that very often the forensic science laboratory or
laboratory system available to the prosecutor in the area did not develop
and maintain a full-service forensic science delivery system. The laboratory system operated in a reactive mode. Such a system carries out
tests and reports results. The attention is focussed narrowly on “items
and tests” rather than on the scientific interpretation of a larger physical
evidence picture. Over this period of time spanned by my observations
the situation has worsened. It needs to be addressed. The interpretation
of scientific data and findings is a scientific problem. It is critically
important that this interpretation be done by scientists. It should not be
left to prosecutors, defense attorneys, or judges to make scientific interpretations. Once appropriate scientists have rendered the scientific interpretation, investigators, members of the legal profession, and the finders
of fact can then use it. Laboratory systems that do not address this need
* Presenting Author
are not supplying a full forensic science service. A void or empty niche
is left. This default by the forensic science service can lead to an even
more disturbing and insidious problem than that just described. It may
lead scientifically naive prosecutors to seek outside expertise from individuals who have established reputations as “good witnesses” but who
lack appropriate scientific credentials. This phenomenon is experiencing
alarmingly rapid growth. The desirability of scientists integrating the
information and rendering overall conclusions is great. However, there
are possible pitfalls. It is important to recognize that certain overarching
conclusions may not call for scientific expertise. There is a danger of scientists wrongly giving common sense conclusions the imprimatur of
science. This must be recognized and avoided. A balance must be struck
between ignoring the need for scientific interpretations and the excesses
of over interpretation. Appropriate interpretation is at least as important
as the laboratory work.
The situations justifying the use of an outside expert should be relatively rare. However, such needs do arise. Some areas of trace evidence
can be very arcane and beyond the experience of many trace evidence
experts. Ideally, when an outside expert is brought into a case, it should
be at the suggestion of the forensic science laboratory system or at least
in consultation with it. If this is not the case, it is cause for concern. This
suggests that there is a problem with the relationship between the laboratory and the prosecutor’s office, that there is something wrong with the
proposed “expert,” or that there is a problem with the laboratory itself.
The problem could be some combination of the three. When such a situation arises, it constitutes a warning sign that should be heeded. The
perils are ineffective case solutions on the one hand, or worse, pseudoscientific testimony and the possibility of a miscarriage of justice on the
other.
Public Laboratories, Private Consultants, Physical Evidence
Interpretation
B40
Forensic Scientists at the Crime Scene:
Lab Directors’ Perspective
Robert C. Shaler, PhD* and Peter A. Pizzola, PhD, New York City
Office of Chief Medical Examiner, 520 1st Avenue, Department of
Forensic Biology, New York, NY 10016
science does not start at the laboratory door. From a laboratory
director’s point of view there is a need for scientific assessment of
physical evidence from the scene until trial.
This presentation will impact the forensic community by helping to
bring into open discussion the need for the scientist at the crime scene.
For too long the crime scene has been viewed as an entity unto itself
that can be properly handled as a separate issue from laboratory
analyses. This is a regrettable abridgement of the scientific continuum
(crime scene to trial testimony) that has existed for a number of reasons
that are primarily bureaucratic or political.
Almost all physical evidence generated in casework is dependent
on the competency of the process used for this purpose. Since this is, in
essence, the foundation for which all the other case examinations are
built, how can it occupy a less important position in the hierarchical
ladder of evidence? The reliability of all analyses that are conducted subsequent to the crime scene can be no more reliable than the work done
at the scene. How important are the things that are not detected at the
crime scene? What impact could these items have on the outcome of a
case? The evaluation or interpretation of all data generated by all the lab
analyses cannot be of any more value than that gleaned from the crime
scene, i.e., the crime scene is the limiting factor in all cases. Simply
stated, the best of laboratory work cannot compensate for a crime scene
inadequately handled.
Historically, police department administrators have decided who
shall be responsible for processing crime scenes or for conducting crime
* Presenting Author
scene reconstructions. Traditionally, this responsibility has been placed
with police officers, acting as evidence technicians, because they are
plentiful in number, are readily supervised in quasi-military fashion, and
have other built-in advantages for scene security and so forth. Much of
this philosophy or approach of utilizing technicians in place of scientists
stems from many naive administrators and even forensic scientists incorrectly believing that technology and science are synonymous. It is a
common misconception that if technological prowess is achieved or
modern gadgetry used that this equates to competent science. It clearly
does not.
The use of a scientific approach in investigating major cases crime
scenes is critical. It has been stated in a noteworthy forensic journal that
the scientific method must be used for crime scene reconstruction.
However, it has also been stated incorrectly, in the same journal article,
that crime scene reconstruction is not science itself, since it is the application of science to matters of law. Forensic science itself is commonly
defined in essentially the same way. Would anyone in the AAFS be
willing to admit that forensic science is not science because it is the
application of science to matters of law? Can non-scientists be allowed
to define forensic science? Can a non-scientist be expected to skillfully
employ the scientific method at crime scenes? How do you become
skilled in the use of the scientific method and its application to crime
scenes? Don’t some cases require follow-up experimentation? How are
experiments designed properly in the crime scene context by untrained
personnel that boast of workshops as the core of their so-called scientific
training? What about the philosophy of science? The questioning of
one’s own findings? Can this be taught effectively in a workshop setting
by non-scientists? How can a non-scientist adequately determine what
scientific examinations need be done in a forensic science lab in a
complex major case.
The above questions will be explored in detail. Several cases will
be presented that illustrate and underscore the above concepts. An ideal
model for the investigation of crime scenes will be proposed and key
recommendations made.
Forensic Scientist, Scientific Assessment, Crime Scene
B41
So Where Do We Go From Here?
Defining Roles for the Forensic
Scientist in Criminal Investigations
Brian J. Gestring, BA, MS*, Pace University, 1 Pace Plaza, Room
Y33C - Forensic Science Program, New York, NY 10038
This presentation will illustrate the changing role of the forensic
scientist in criminal investigations and discuss how the forensic scientist
can influence this change.
This presentation will draw attention to the changing role of the
forensic scientist in criminal investigations and illustrate how we can
influence our future.
Never before has forensic science enjoyed the exposure that it currently receives. Dramatic depictions and televised trials have increased
the public’s awareness of the value of physical evidence. Increasingly,
juries influenced by these forces are expecting more from criminal
investigations, often to what seems like unrealistic extremes.
Yet if we look critically, are these extremes so unrealistic? Who
would have imagined the level of discrimination that we now regularly
obtain with biological evidence? The significance of this advance cannot
be understated. Nor can the jarring effect that this advance has had on
the rest of the field.
Forensic science has changed in response to this new individualizing potential. Data basing systems have converted DNA results, formerly only useful once a suspect had been identified, to a valuable investigative tool. Unfortunately the change was not always a positive one.
Juggling already limited budgets, laboratory administrators have been
forced to reevaluate their priorities. Some even choosing to forgo
43
conventional trace analysis altogether.
The nature of the scientist performing the analysis has also
changed. As tools of our trade become more complex, it is becoming
increasingly difficult to resist the slippery slope toward specialization.
Some would argue that this natural phenomenon represents the evolution
of the field. If left unchecked, this change would be better classified as
erosion. This is not to say that the specialist does not play an essential
part in contemporary analysis, only that the specialist must be used in
conjunction with a generalist to maintain the necessary perspective.
This combined scientific approach will allow us to lay the root-system
which will ultimately change how forensic science is practiced.
Nowhere is the generalist viewpoint more important than at the
inception of the investigation. Here at the scene of the crime, critical
information can be recognized, documented, and collected. Failure at
this stage will prevent any subsequent laboratory analysis, no matter
how sophisticated. Even the staunchest opponents will concede this
point. Yet still, forensic scientists have gladly abdicated their role at the
crime scene.
Our investigatory process is fundamentally flawed when non-scientists are charged with posing the scientific questions that will dictate
all subsequent analysis. This error is further compounded when laboratories are relegated to solely a reactive role. Much like the proverbial
“black box,” investigators and prosecuting attorneys will indicate the
testing they require as a case progresses and the laboratory will just be a
mythical place where the answer is generated.
While this is our present, it does not have to be our future. More so
now then ever, we have the ability to change how science will be practiced in criminal investigations. What better way can forensic scientists
serve as a counterbalancing weight between truth and justice than
through charting a team approach, and by ensuring that investigator and
scientist work side-by-side with scientific assessment of the evidence at
every stage of the process?
This paper will focus on current realities of the situation and outline
a systematic approach for implementing these much-needed changes to
our role as forensic scientist in criminal investigations.
Scientific Assessment, Crime Scene, Future Role
B42
Development of a Portable X-Ray
Fluorescence Instrument for
Forensic Applications
Jeffrey S. Schweitzer, PhD*, University of Connecticut, Department of
Physics, Unit 3046, 2152 Hillside Road, Storrs, CT 06269-3046; Jacob
Trombka, PhD, Samuel Floyd, MS, and Yossi Eisen, PhD, Goddard
Space Flight Center, Greenbelt, MD 20850; Carl Selavka, PhD,
Massachusetts State Police Crime Laboratory, Sudbury, MA 20850;
Gerald Zeosky, MPA, New York State Police Forensic Center, Albany,
NY 18015; W. Mark Dale, New York City Police Forensic Laboratory,
Queens, NY 18012; Norman Gahn, JD, Office of the District Attorney,
Milwaukee, WI 44106; Elyse Gottschang, MFS, Goddard Space Flight
Center, Greenbelt, MD 20850; James Marie, MS, University of
Connecticut, Storrs, CT 06269; Raymond Kimble III, BS, ACS Defense,
Inc., Washington, DC 20850
After attending this presentation, participants will understand the
critical design features and operational parameters of a new, portable xray instrument for assisting in the recognition of trace evidence for crime
scene and laboratory applications.
This presentation will impact the forensic community by: 1)
describing the development of a portable x-ray fluorescence instrument;
2) demonstrating the operational performance of the instrument for trace
residua of forensic importance, including primer residue, blood, and
semen; and 3) discussing the future use of this instrument at crime
scenes or in the laboratory to develop investigative leads by assisting in
the recognition and recovery of such trace evidence.
44
A ruggedized x-ray fluorescence instrument has been designed to
investigate trace element content at crime scenes. The initial focus is to
identify possible materials of interest such as gunshot residue (GSR) and
bodily fluids. The instrument has been designed to be part of a system to
aid crime scene investigation and transmit the data to locations requiring
it. This portable instrument was designed to meet the obvious constraints
of weight, battery operation, and ruggedness. A number of special design
features, however, were needed to allow for the detection of microgram
quantities of the trace elements of interest. The instrument makes use of
a custom-designed x-ray generator and a new type of Shottky-barrier
cadmium telluride x-ray detector.
This instrument is part of a teleforensics program jointly funded by
NIJ and NASA. This collaboration seeks to develop cost-effective
instrumentation based on technology developed for the space program to
benefit crime scene investigation, and to develop advanced instrumentation for planetary missions for NASA. A critical factor at crime scenes
is the collection of evidence for analysis at forensic laboratories. The
friable nature of evidence requires rapid recognition, to avoid losing the
probative information contained therein. Some evidence is invisible to
normal investigation techniques, either because it involves trace quantities not visible to any investigation technique, or because it is covered
or hidden from view. Many types of potential evidence can be indicated
by crime scene detection through in situ trace element analysis.
We show data that supports possible use of an x-ray fluorescence
instrument through detection of gunshot residue, blood (through the
detection of the iron in hemoglobin) and semen (through the detection of
zinc protoporphorin). To detect the low levels of trace element concentrations, advanced technology has been incorporated including the
unique x-ray generator and a recently developed Shottky cadmium telluride x-ray detector. The design of the internal structure of the
instrument minimizes the background due to coherent scattering. It was
also necessary to select the x-ray tube anode material to ensure that the
tube’s x-ray line production does not interfere with the detection of elements of forensic interest.
Results of experiments to date realistically characterize the
expected performance of the instrument for detecting trace element concentrations that are useful for investigating crime scenes and in laboratory applications.
X-Ray, Trace Evidence, Criminalistics
B43
Visualization of Bloody Fingerprints
at Violent Crime Scenes
Ellen J. Bishop, BS*, Toby L. Wolson, MS, and Gilberto Tamez, MiamiDade Police Department Crime Laboratory Bureau, Forensic Biology
Section, 9105 NW 25th Street, Miami, FL 33172
After attending this presentation, attendees will have a better understanding of the causes of bloody fingerprints that are frequently
observed at violent crime scenes and the significance of the visualized
bloody fingerprint.
This presentation will provide the analyst with a better understanding of how bloody fingerprints may be visualized at violent crime
scenes. It will assist in the interpretation and presentation of bloody fingerprints when doing bloodstain pattern analysis evaluations and expert
witness testimony.
Fingerprints were deposited on a variety of substrates that may be
encountered in the home or workplace. The substrates that were used for
the research were pieces of glazed ceramic tile, glass, melamine bookshelf, wood (pine and poplar), acrylic plastic, residential vinyl floor tile,
commercial vinyl floor tile, and galvanized sheet metal. The substrates
were cleaned prior to use and several of the pieces of wood and galvanized sheet metal were painted using either oil-based glossy, oil-based
matte, latex glossy, or latex matte paint. Each of the painted substrates
was also tested in an unpainted condition. After preparation of the
* Presenting Author
substrates, fingerprints were deposited on them using either light or
heavy pressure. The fingerprints were made by coating a finger with
motor oil, WD-40® spray lubricant, floor wax, skin oils, hair oils, or silicone lubricant. Fresh human blood mixed with citrate phosphate dextrose anticoagulent obtained from the South Florida Blood Bank was
then dripped, spattered, or flowed over the fingerprints. The interaction
of the blood with the fingerprint was recorded; fingerprints that became
visible upon interaction with the blood were photographically documented. The photographs of the bloody fingerprints were examined by
a Latent Print Examiner to determine if the fingerprint was a positive or
negative image and if it was of comparison value.
A second study examined the visualization of bloody fingerprints as
the result of touching a blood drip. Blood was deposited on the substrates in volumes of 50µl. A finger was then pressed into a pool of
blood at 0, 15, 30, 45, and 60 minutes. The results were photographed
and examined by a Latent Print Examiner.
The third study examined bloody fingerprints that resulted when
the substrate was touched by a finger that had been coated with blood.
The results of the experiment were photographed and examined by a
Latent Print Examiner.
The substrate characteristics, as well as the method used to deposit
the blood on the substrate, affected the formation of visible bloody fingerprints. The less porous, smooth, polished, and/or glossy substrates
resulted in the visualization of a higher number of bloody fingerprints.
Flowing or dripping the blood onto the substrates resulted in bloody fingerprint images more often than the test substrates that were spattered
with blood. The fingerprints made from oils interacted more often with
the blood to leave a visible fingerprint than the fingerprints made from
silicone lubricant or wax. These visible bloody fingerprints were negative or reversed images. The fingerprints became visible because the
blood flowed into the areas that were not oily. These non-oily areas correspond to the furrows of the fingerprint and the oily areas represent the
ridge pattern of the fingerprint. All of the bloody fingerprints that were
visualized by interaction with the human blood during the first and
second studies were negative images and none were of comparative
value. The bloody fingerprints from the third study were all positive
images and were of comparative value.
Bloody fingerprints are frequently identified at violent crime
scenes. It is frequently argued that the fingerprint was there prior to the
bloodshed event and that the blood interacted with the fingerprint to
make it visible. Another argument is that the defendant entered the
crime scene after the bloodshed event and accidentally touched a pool of
semi-dried blood, leaving a bloody fingerprint in its place. This research
demonstrates that it is difficult to obtain bloody fingerprints as a result
of a pre-existing fingerprint interacting with blood being deposited on it
or because a finger was pressed into a pool of semi-dry blood. When
bloody fingerprints are visualized due to these interactions, the resulting
fingerprint is a negative image of no comparative value. The only mechanism that will consistently produce a positive image of comparative
value is when a finger coated with blood contacts a clean substrate and
transfers the blood onto that substrate.
The research provides the analyst with a better understanding of
how bloody fingerprints may be visualized at violent crime scenes. It
will assist in the interpretation and presentation of bloody fingerprints
when doing bloodstain pattern analysis evaluations and expert witness
testimony.
Bloodstain Pattern Analysis, Bloody Fingerprints, Latent Print
Examination
* Presenting Author
B44
Burnin’ Down the House: Predictive
Cadaver Models in Structural Fires
Elayne J. Pope, MA*, University of Arkansas, 330 Old Main,
Fayetteville, AR 72701; Robert Fenton, Hickory Creek Volunteer
Fire Department, Lowell, AR 72745; O’ Brian C. Smith, MD,
Regional Forensic Center, 1060 Madison Avenue, Memphis, TN 38104;
Jody Fenton, BA, University of Arkansas, 330 Old Main, Fayetteville,
AR 72701
After attending this presentation, authors will have: 1) demonstrated benefits of collaborative research with Arson Investigation,
Anthropology, and Pathology, 2) established how burn patterns are influenced by structural conditions within the fire, 3) illustrated scene reconstruction using patterns and condition of burned human remains, and 4)
interpreted body patterns as part of the scene.
This presentation demonstrate the importance of how identification
and recovery of fragmentary bone are valuable specimens when reconstructed and provide specialized information about personal identification or preexisting trauma of human remains.
Discovery of human remains in structural or vehicular fires presents many challenges to the investigative process. Traditionally, expedient removal of human fatalities precedes the comprehensive structural
analysis of the scene. Ideally, the independent examinations of burn patterns at the scene and on the body should correlate with reconstructed
events surrounding the fire. However divorcing analyses of the two
types of physical evidence invites the potential for information loss
when examined in isolation. This presentation demonstrates how a contextual analysis of human remains in situ strengthens the fire death
investigation as burn patterns of the body are interpreted within the
unique conditions of the scene.
Similar to charred patterns on walls, burn patterns on the body
readily disclose information about its orientation to heat, position within
the fire, exposure, and presence of preexisting trauma. When human
remains are examined as an integral part of the original fire scene this
context additionally provides relative proximity to exits, entrapment or
protection by collapsed debris, and spatial orientation within the
structure. However, in most fire death cases human remains are often
removed in haste, leaving valuable fragments of soft tissue, bones, or
teeth camouflaged among debris as overlooked and permanently lost
specimens. An awareness of how the body burns in response to different
spatial and structural conditions aids in reconstruction since complete
recovery of these fragmentary remains is not always accomplished.
Collaboration with the Hickory Creek Volunteer Fire Department,
Anthropology Department, and Shelby County Medical Examiner conducted field experiments creating predictive cadaver models within
structural fires to improve reconstruction techniques for arson investigation. Actual research using unembalmed human bodies from
anatomical gift donations and condemned residential structures were the
primary research materials. Bodies were strategically placed in known
locations and positions within different types of structural settings.
Variable selected conditions include direct placement upon carpeted
concrete foundation slab, elevated floors over crawlspace or basement,
and placed upon the second level of a multiple story house to examine
burn patterns and spatial distribution of associated fragmentary remains
for each situation during the post burn recovery phase.
Fully ignited structures were documented for the entire duration:
recording known conditions of materials, times, and temperatures until
natural extinguishment. Remains of the structure and body were left to
smolder and excavated the following day. Collaboration of archaeological and arson investigative techniques were employed to interpret
the contextual position, condition, and spatial patterns of human remains
and associated fragmentary pieces camouflaged among debris. In most
cases fragmentary remains of extremely incinerated bodies fell within
close proximity beneath the resting position. Relative differences in
45
ventilation access and variable height correlated with the distribution
and extent of burning for each body. Fragmentary remains lying directly
on the concrete foundation were larger, easily identified, and predictably
positioned compared to conditions with increased distances between the
body’s vertical position and ground level. For the latter combinations of
vertical descent, impact with architecture (flooring and joists), and
dimensional collapse of debris contributed to morphological differences
for surviving remains of elevated cadavers, producing smaller fragments
and wider dispersal area.
The use of different residential structural types generates predictive
models to help arson investigators recognize how the body burns in
response to environmental conditions and effectively identify expected
distributional patterns of fragmentary human remains during recovery.
In addition to examining physical evidence of the scene, burn patterns of
soft tissue and bone are equally valuable tools used to reconstruct events
of the fire. This presentation stresses how identification and recovery of
fragmentary bone are valuable specimens when reconstructed and
provide specialized information about personal identification or preexisting trauma of human remains. Results from these experimental burns
demonstrate the fruits of collaborative multidisciplinary field research
and offer new information for effectively investigating fire fatalities.
Fire Investigation, Burned Bodies, Burned Bone
B45
Self-Cleaning Window Glass:
Breakage Transfer Process Validation
and Subclass/Brand Characterization
Corrie D. Maggay, MFS*and Robert D. Blackledge, MS, Naval
Criminal Investigative Service, Regional Forensic Laboratory, 3405
Welles Street, Suite 3, San Diego, CA 92136-5018; Faye Springer, BS,
Sacramento County District Attorney Crime Laboratory, 4800
Broadway, Suite 200, Sacramento, CA 95820
Via high-speed videos filmed at 1000 frames/sec. attendees will be
witness to the process of window glass breakage and to the forces producing numerous fragments going back towards the breaker. The goals
of this presentation are to graphically validate this process of glass
fragment transfer, to qualitatively assess the likelihood that transferred
particles tend to originate from the glass surface towards the breaker, and
to present results from tests on recovered glass fragments that permit
brand identification.
This presentation will study a breaking and entering entrance commonly gained through breaking a window. Although glass does not
break back onto the perpetrator, explaining it to a jury can be a trying
task. A jury is composed of twelve people, many of whom do not have
a background in science. Forensic scientists’ ability to explain work and
examinations to a jury takes great precedence over the work itself, for it
is the jury who is judging the defendant. 90% of all people are visual
learners. The videos shown in this presentation will be available to the
forensic community with hopes that they will one day be of some use in
the courtroom.
Via high-speed videos filmed at 1000 frames/sec. attendees will be
witness to the process of window glass breakage and to the forces producing numerous fragments going back towards the breaker. The goals
of this presentation are to graphically validate this process of glass
fragment transfer, to qualitatively assess the likelihood that transferred
particles tend to originate from the glass surface towards the breaker, and
to present results from tests on recovered glass fragments that permit
brand identification.
Self-cleaning window glass, a new type of glass intended solely for
exterior use, was introduced to the AAFS audience in a presentation by
one of the authors last year. One side only of these panes (the exterior
facing side) is rich in titanium dioxide (TiO2), the catalytic ingredient
that in concert with the sun’s ultraviolet rays acts as an oxidizing agent
46
and breaks down organic dirt and grime. Since only glass at or very near
the exterior surface is rich in TiO2 it is possible to not only discriminate
this new subclass from ordinary window glass, it is also possible to
identify tiny fragments originating from this surface.
Window breaking tests were conducted at the U.S. Army Proving
Ground, Yuma, Arizona, utilizing available high-speed cameras. Under
identical conditions, 1’ x 1 ‘ x ¼” glass panes mounted at approximately
shoulder height were broken by swinging a crowbar. Panes broken
included ordinary window glass, and the two available self-cleaning
brands, Activ™ glass by Pilkington and SunClean® glass by Pittsburgh
Plate Glass. The self-cleaning panes were mounted with the treated side
towards the breaker.
With the exception of different colored hoods, the breaker wore
identical outfits for each glass-breaking event. Subsequent to each
breakage the breaker moved over to an area where clean butcher paper
was laid down, and the breaker’s outer clothing (hood, gloves, sweater,
sweat pants, and sneakers) were separately removed and packaged as
evidence. Later, these items were searched for glass fragments. Glass
fragment recovery data and results of characterization examinations
(fragments originating from the surface towards the breaker, from the
middle, and from the side opposite the breaker) will be presented.
Trace Evidence, Glass Breakage and Analysis, Self-Cleaning
Window Glass
B46
A Validation Study of Physical
Associations of Duct Tape Ends
Maureen J. Bradley, PhD*, Roger L. Keagy, BS, Preston C. Lowe, MS,
Michael P. Rickenbach, PhD, Diana M. Wright, PhD, and Marc A.
LeBeau, MS, Federal Bureau of Investigation, Laboratory Division,
2501 Investigation Parkway, Quantico, VA 22135
By attending this presentation, attendees will learn how a study to
evaluate the validity of conducting physical associations (endmatches)we designed and administered. Although we looked at duct
tape, a common consumer item submitted to forensic science laboratories, the study can be used as a model to evaluate other materials (such
as glass, wood, metals, paper) submitted to forensic laboratories. The
results of the study can be used by other forensic science laboratories
that conduct end-match examinations on tape to support their test results
in court challenges.
This presentation will impact the forensic community by demonstrating that although tape end-match examinations have been conducted
in forensic science laboratories for decades, there are no published validation studies on this technique. Presentation at this conference will be
the first opportunity to inform the forensic community of the results of
the study. The design and administration of the test will be presented so
other laboratories may use the study as a model to evaluate other types
of materials commonly submitted to forensic science laboratories for
physical association (end match) examinations such as metal, wood,
glass, etc.
Although tape end-match examinations have been conducted in
forensic science laboratories for decades, there are no published validation studies on this technique. In the era of Frye and Daubert where a
forensic examiner must demonstrate to the court the scientific soundness
of their examinations, we undertook a study to address this concern.
The purpose of the study was to determine the validity of conducting physical association (end match) exams on duct tape evidence
and to evaluate the error rate associated with such an examination. The
study was also designed to address the following: the number of associations/end matches identified correctly and incorrectly; the ability to
identify associations/end matches for both torn and cut pieces of duct
tape; and whether the grade of tape influences the ability to identify
associations/end matches.
* Presenting Author
Three rolls of duct tape were used, representing different manufacturers and grades of tape. Each participant received five sample sets,
three sets with hand-torn ends and two sets with scissor cut ends.
The sample sets were prepared as follows: ten (10) strips of each
tape were either cut or torn and adhered sequentially onto a plastic
surface. Each strip of tape was randomly labeled alphabetically and then
numbered sequentially (1-10) on the plastic surface to which the tape
was adhered. Each test set was digitally documented and the order of
tape within each test was verified by a second person. The pieces of tape
were then separated from one another by cutting the plastic substrate.
The substrate was cut in such a way to remove the sequential number
and so that end-matching of the substrate was not possible. For each test
set, the ten (10) strips of tape were shuffled together and three (3) strips
were randomly removed. The three (3) randomly removed strips were
placed into a properly labeled plastic bag. The remaining seven (7)
strips were placed into individual plastic bags and labeled appropriately.
A total of 20 sample sets were administered to four analysts with
directions to evaluate whether or not physical associations (end matches)
existed among the strips of tape in each set following the standard operating procedure.
The results of the initial study were evaluated by the test administrator. In cases where there was a failure to report a physical association
by the original test participant, the test set was evaluated independently
by the three remaining test participants and each rendered their opinion.
The results of this study will be presented in detail addressing each
of the aforementioned objectives. Discussion will include the number of
correctly identified tape end-matches, whether the manner in which the
tape is separated (hand-torn versus cut with scissors) and whether the
grade of tape has an effect on the ability to conduct these examinations.
Duct Tape, End-Match, Validation Study
B47
Frequency of Occurrence
Data for Textile Fibers
Harold A. Deadman, PhD* and Tara A. Scully, BS, George Washington
University, 2036 H Street, NW, Washington, DC 20036
After attending this presentation, attendees will learn that most
textile fibers have considerable evidential value as associative evidence
This presentation will demonstrate the importance of considering
the evidential value and meaning of associative evidence using textile
fibers as an example.
The question, “What does the evidence mean?” is obviously an
important question in a criminal trial. Determining the value and
meaning of evidence is one of the most important factors that go into
forensic analysis. However, it is often one of the most difficult questions
for the forensic scientist to answer objectively and to provide objective
support for that answer in the courtroom. Because of this, the question
is often ignored in laboratory reports and in testimony at a trial.
Assessing and presenting evidential value is especially difficult with
respect to trace evidence. Evidential value of a particular type of evidence can vary tremendously depending on the specific evidence
available in a case. Forensic scientists must consider the evidential
value and be able to present and support their assessment in the
courtroom as a jury will not have the knowledge necessary to correctly
evaluate the evidence.
Assessing evidential value generally occurs after matching evidence from two objects or locations has been found to exist, resulting in
an association. For example, textile fibers recovered from a homicide
victim’s body are matched to a carpet in a suspect’s residence linking the
* Presenting Author
victim to the suspect’s residence. However, since it is possible that these
carpet fibers came from a similar but different carpet, it could be a coincidental match and, therefore, the probability of a coincidental match
must be addressed. To simply say that the fibers on the body are consistent with originating from the suspect’s carpet does not address the
evidential value and is likely to downplay the significance of the fiber
evidence.
There are many factors that go into the assessment of the evidential
value of fiber evidence. One of the most valuable pieces of information
the fiber examiner could use in assessing evidential value is the frequency of occurrence of the probative fibers in textile materials or in
debris from textile materials. This type of information may be difficult
if not impossible to determine for a fiber type involved in a specific case,
however, it is possible to determine an average or relative frequency of
occurrence for particular fiber types, which can then be equated with the
fibers of interest in a particular case.
Over the past 20 years there have been many “target fiber” and
“population studies” that have addressed the issue of the frequency of
occurrence of specific fiber types in clothing and in debris collected
from various locations. These studies have shown that, although there
are some very common fibers which would have little value as evidence,
a fiber that is not one of the very common types will have a very small
frequency of occurrence. Most of these studies have been conducted
outside the United States, primarily in Great Britain, Germany and
Australia. Although these studies would likely apply to fiber analysis in
the United States, it would be helpful to conduct similar studies in the
United States.
This project is designed to provide data useful in assessing the evidential value of fiber evidence by providing information on the frequency of occurrence of particular fiber types. It consists of three component parts: first fibers have been collected from items of clothing. The
clothing sampled came from university students as well as from office
staff wearing both casual and business dress. These fibers are being
classified initially based on microscopical characteristics such as color,
fiber type and diameter. This data can be used to address the frequency
of occurrence of different fiber types in clothing. A second component
of the project is the collection of fibers from 50 seats and 50 seat backs
located in a University auditorium. After using tape to collect the fibers,
approximately 10% of the recovered fibers have been mounted and
searched until 40 dissimilar fiber types have been identified from each
taping. A total of 4000 fiber types will have been identified and classified in approximately 1000 possible classes at the completion of the
project. Fibers within a class will then be compared. This data will also
address the frequency of occurrence of fiber types from clothing and
debris from clothing that has been transferred to the seats and seat backs.
A third component of this research is to search the debris from the auditorium seats and seat backs for several fiber types that are present in the
composition of three sweaters. The three sweaters selected as targets are
various shades of red and at least 50 red fibers were recovered from each
taping. The more than 5000 red fibers from the auditorium seats are
being compared with the rayon, acrylic, polyester, nylon and cotton
fibers in the composition of the three sweaters.
The data obtained thus far support the conclusions that have been
reported in the literature in studies outside the United States. While
there are some common fiber types, there is tremendous variety in the
world of textile fibers. As long as a fiber is not one of the common fiber
types, the probability of finding a particular fiber type in a particular
location or in the composition of a particular item of clothing by chance
is very small. This is particularly true with respect to man-made fibers
and especially with colored man-made fibers.
Textile Fiber Evidence, Evidential Value, Fiber Frequency of
Occurrence
47
B48
Application of Principal Components
Analysis in the Individualization of
Gasolines by GC/MS
Jeremy P Wintz, BS* and J. Graham Rankin, PhD, Forensic Science
Program, Marshall University, 1401 Forensic Science Drive,
Huntington, WV 25701
After attending this presentation, attendees will understand the usefulness of multivariate statistics to forensic applications in particular
GCMS.
This presentation will impact the forensic community by demonstrating the usefulness of multivariate statistical analysis to large data
sets especially in comparisons between GCMS data in gasoline individualization.
Gasoline is an accelerant frequently used by arsonists.
Identification of gasoline in fire debris is relatively easy even when
greatly evaporated. When a suspect is apprehended with gasoline
residue on his clothes or a gasoline can in his vehicle, the question arises
if the gasoline residue from the fire debris can be matched with that
found with the suspect. This has proven to be a more difficult challenge.
Further recent legal challenges to comparison evidence have stressed the
necessity of establishing a statistical probability for that match.
Julia Dolan (ATF National Research Lab, Ammendale, MD), at the
Atlanta meeting of AAFS, presented a high-resolution GCMS method
for comparing gasolines based on 20 sequential area ratios of 34 target
compounds from 3-methylpentane through the 1-methylnaphthalene.
Her data set included 36 different gasolines, including both regular and
premium, mostly from around the Washington, DC area. In addition to
neat gasolines, 25% and 50% evaporated samples were analyzed.
We have applied principal components analysis (PCA) to her data
set as well as our own collection of over 60 gasoline samples from across
the US that we analyzed by her method. Most of the sequential ratios
are reproducible in triplicate analysis to less than 5% relative standard
deviation. The ratios show little, if any, change between un-evaporated
and 50%-evaporated gasoline. However, based on our PCA results, of
the 20 ratios, only a few show a significant contribution to the betweengasoline variation thus being able to distinguish one gasoline from
another. Because some of these distinguishing ratios are in the light
petroleum distillate range, they may be lost in higher evaporated
gasolines.
Gasoline Analysis, Principal Components Analysis, Fire Debris
B49
Determination of Hand Deposited GSR
Obtained From Shootings Carried Out
With a Pistol and Cartridges Produced
by TMCF Using SEM/EDS Method
Ismail Cakir, PhD*, Council of Forensic Medicine, Turkey, Adli Týp
Kurumu, Cerrahpaþa, Istanbul, 34246, Turkey; H. Bülent Üner, PhD
and Salih Cengiz, PhD, Institute of Forensic Sciences, Istanbul
University, Adli Týp Enstitüsü, Cerrahpaþa, Istanbul, 34301
Shooter identification is of importance in criminal firearm cases in
order to reveal the manner of death or injury (murder, suicide, accident).
Scientifically, shooter identification is possible by analyzing the gunshot
residues (GSR) obtained from the suspects’ hands.
48
When a weapon is fired, GSR are discharged at high velocity from
the muzzle and are expelled from ejection port. Some of the GSR that
are expelled from the ejection port are deposited on the firing hand.
There is no scientific study about the analyses of GSR on firing
hands of Turkish made ammunitions. For this purpose experimental
shootings were carried out using 9mm Parebellum type cartridges, all
produced by Turkish Machining and Chemistry Foundation (TMCF) and
a Atmaca 53 pistol performing one, two, three and four shots with the
right hand.
Firing hands were sampled by using aluminum stubs coated with
double-sided adhesive. Shooter’s hands were washed and dried after
each test firing.
SEM/EDS analyses for GSR particles were carried out using a
JEOL 5600LV scanning electron microscope equipped with a LINKISIS 300 X-ray analyzer.
The results obtained in this study indicate that elemental composition of the GSR of the studied cartridges were found compatible with
the elemental composition of the original chemicals of them. The GSR
results showed the following elemental distribution to TMCF cartridges:
unique particles (PbSbBa, SbBa), indicative particles (PbSb, PbBa,
Pb,CuZn, CuZn, Cu, Zn, Fe, Pb).
Gu Shot Residue, SEM/EDS, Turkish Machinery and Chemistry
Foundation
B50
Comparison of DNA Stability Stored
on Treated and Untreated Papers
Frank Igoe, PhD, Betsy Moran, PhD*, Renate Karle, MS, and Martin
Smith, PhD, Whatman, Inc., 9 Bridewell Place, Clifton, NJ 07014
After attending this presentation, attendees will have knowledge of
DNA stability over time on FTA treated paper v untreated filter paper.
The attendee will make an informed decision of how to store DNA for
long periods of time.
humanity by providing data on the various options of long term DNA
storage.
In forensics, the integrity of the sample DNA is crucial for successful analysis. Often, DNA is collected by applying samples (blood,
saliva, tissue, etc…) to chemically treated papers such as FTA® or
untreated papers such as 3MM or 903®. These papers are a convenient
medium for collecting DNA: cells are lysed on contact with the paper
and DNA is bound to the matrix. Sample DNA on the treated paper is
stabilized and protected against degradation. Samples may be kept at
room temperature for long periods of time and are easily made ready for
analysis by PCR based amplification. Up to this time, there has been no
systematic analysis of the integrity of DNA samples stored on the treated
vs. untreated papers.
Here we present data showing that FTA®-treated paper provides
greater protection of genomic DNA from degradation than untreated
papers. This increased protection was consistent and was observed
within hours of application and up to 180 days after sample application.
This protection is seen in samples whether stored at room temperature,
at -20 °C or at 37 °C with high humidity. These results demonstrate that
DNA stored on FTA is safe for long-term storage thus increasing the
probability of obtaining reliable testing results.
DNA Stability, Treated Filters, FTA
* Presenting Author
B51
Evaluation of the Least Square
Deconvolution Approach in
Interpreting DNA Mixtures
Suzanne D. Shaffer, MS* and Christina Capt, BS, University of North
Texas Health Science Center at Fort Worth, 3500 Camp Bowie Blvd,
EAD 1-348, Fort Worth, TX 76107; Tsewei Wang, PhD, Department of
Chemical Engineering, University of Tennessee, 419 Dougherty Hall,
Knoxville, TN 37996-2200; John V. Planz, PhD, University of North
Texas Health Science Center at Fort Worth, 3500 Camp Bowie Blvd,
Fort Worth, TX 76107
The goal of this presentation is to present the use of mathematical
models to resolve DNA Short Tandem Repeat profiles.
This presentation will provide a mechanism for interpreting DNA
mixtures in forensic casework including but not limited to sexual assault
evidence, trace or low copy number samples, product of conception
cases, and commingled remains from mass disasters.
Introduction: DNA samples containing mixed profiles are often
encountered in forensic DNA casework, stemming most often from
sexual assault cases. Characteristic peak height imbalances and disproportionate intensity seen in short tandem repeat (STR) analysis indicate
that DNA contributions may originate from different individuals. The
U.S. Supreme Court established under Daubert v. Merrell Dow
Pharmaceuticals, Inc. that scientists are required to show the reliability,
reproducibility and validity of their scientific results. Current mixture
resolution methods do not rely on all available empirical data. Mixture
statistics such as the probability of exclusion fail to differentiate between
the victim and suspect profiles and do not take into account the ratio of
mixtures. Other statistical measures such as likelihood ratios take into
account major and minor peak levels but fail to provide a measure of differing mass ratios. Least Square Deconvolution (LSD) algorithms
strengthen the reliability, reproducibility and validity of mixture interpretations required by Daubert standards by providing a systematic
mathematical approach to resolving mixtures. All available data generated in electropherograms are evaluated. LSD software computes a
mass proportion for each two contributing genotypes by comparing their
relative peak height and/or area measurements assuming 1) relative mass
ratio is approximately preserved during PCR amplification across all
loci and all alleles within a locus, and 2) allele peak heights and areas are
proportional to its relative DNA mass. The software takes into consideration all possible combinations of the two contributing genotypes and
computes the corresponding fitting errors. The higher mass profile is
normalized against the lower mass proportion to calculate the mass ratio.
The profile with the smallest fitting error is determined to be the most
likely genotype scenario. Hypothesis/Methods: Evaluate the effectiveness and precision of the LSD software. Different male to female
DNA ratios were PCR amplified and separated by capillary electrophoresis. Corresponding peak height and area data were entered into
the LSD Software. In addition, different PCR and electrophoresis parameters were examined. Results: Preliminary results indicate LSD
software can separate profiles between 10:90 and 30:70 mixture ratios.
Adjusting PCR and electrophoresis parameters enhances ability to separate profiles at a mixture ratio of 5:95. Conclusion: LSD could prove
beneficial to the forensic community in the future by evaluating mass
ratios in mixture samples. In time this analysis system may be used to
strengthen statistical measurements for mixture evidence to satisfy
Daubert standards.
Least Square Deconvolution, DNA Mixtures, Short Tandem
Repeats
* Presenting Author
B52
Evaluating the False Parentage Rate
and CPI Cut-off of CODIS 13 STR
for Seven Populations
Chang En Pu, MS*, Meng-Yi Chen, MS, Lim-Ming Meng, MS, and
Fang-Chin Wu, BS, Ministry Justice Investigation Bureau, PO BOX
3562, No 74, Chun-Hua Road, Hsin-Tien, Taipei 231, Taiwan; Adrian
Linacre, PhD, Forensic Science Unit, Department of Pure and Applied
Chemistry, University of Strathclyde, Glasgow, UK, 204 George Street,
Glasgow, Scotland G1 1XW, United Kingdom
After attending this presentation, attendees will be able to recognize
that false parentage determination after DNA test is possible.
humanity by demonstrating the cutoff value of CPI for parentage could
be established to avoid false parentage, especially important for unidentified body recognition on immigration blood relative testing.
We report on the use of STR typing for the genetic linkage of
unidentified human remains and the problems associated with false
paternity results. STR loci are chosen and used on based upon their
power of discrimination and ability to multiplex with other STR loci.
Most of the STR loci used in forensic science both for criminal and civil
investigations. The CODIS 13 core STR systems has an average power
of paternity exclusion (PE, to exclude a random man) larger than
0.9999(in trio cases) in many populations. In the identification of
unidentified bodies only one relative (either of parents or either of
son/daughter) is available for testing. This results in duo cases for
parentage building. When allele sharing is found in all the 13 loci, the
probability of parentage could be determined preliminarily. However it
is hard to avoid a false confirmation of the alleged father or false identification of the unidentified bodies. In Taiwan, the national unidentified
bodies CODIS 13 STR database has approximately 680 bodies and 200
families. Sometimes a body matches to more than one individuals (from
different families) and cases with extremely low Cumulated Paternity
Index (CPI) were found. It is necessary to evaluate the false parentage
rate and set a cut-off value of CPI for avoiding false determination of
parentage.
The CODIS 13 population data of 177 African American, 194 US
Caucasian, 202 Southwestern Hispanic, 153 Bahamian, 157 Jamaican,
76 Trinidadian and 1,000 Chinese in Taiwan was collected from published websites of FBI USA or by authors. The cumulated power of
exclusion (PE) for duo for African America = 98.31%, US Caucasian =
98.23%, Southwestern Hispanic = 97.97%, Bahamian = 98.48%,
Jamaican = 98.32%, Trinidadian = 98.61% and Chinese in Taiwan =
98.13%. The data showed that about 1.4 % to 2% of random men could
not be excluded from being an alleged father for the studied populations.
A matching test model was designed to evaluate the practically possible
false parentage rate.
All the collected individuals were paired resulting in 15,576,
18,721, 20,301, 11,628, 12,246, 2,850, and 499,500 pairs for each population respectively. Each pair was checked for allele sharing locus by
locus and the CPI for those pairs with 13 loci sharing was also calculated. Microsoft Excel Macros controlled by a program written by the
authors were used to handle the comparison and calculation. CPI calculations were based on the distribution frequencies for the respective populations. These were 8(0.0514%), 10(0.0534%), 9(0.0443%),
3(0.0258%), 7(0.0572%), 3(0.1053%) and 250(0.0501%) pairs found
with 13 allele sharing loci for the seven populations. False parentage
was noted when the CPI for pairs ranged from 1.76 to 1,950,430. These
were 4.48% CPI lower than 100, and 78.27 % lower than 1,000. If the
suitable CPI cut-off value were used such as CPI=1,000, there would be
less of false parentage.
Though we could reduce the false positive rate by increasing the
cut-off value, the false exclusion rate (a real father excluded as a random
man) would be increased accordingly. In this study many duo paternity
49
cases in Taiwan were found with CPI lower than 1,000 when only
CODIS 13 STRs were typed. If CPI=1,000 was suggested as the cut-off
value these low CPI cases could be classified as false parentage cases.
More real paternity cases have to be observed to set an optimum range
of value for CPI cut-off, or more STR systems are required when
matches were found with low CPI. This is especially the case for mass
and open matching operation of STR database for the unidentified
bodies. If less than 13 STR typing were obtained from the unknown
skeleton owing to degradation of DNA, the CPI would be much lower,
and the risk of false identification would be much higher. In such cases
mtDNA sequencing and other investigating techniques should be added
to enhance the discrimination.
Short Tandem Repeat, CODIS 13, Parentage Test
B53
Macro-to-Micro Interfacing of a
Swab Receptacle With a Microchip
for Total DNA Analysis
Benjamin R. Schroeder, BS*, Jerome P. Ferrance, PhD, and James P.
Landers, PhD, Department of Chemistry, University of Virginia,
McCormick Road, Charlottesville, VA 22904
After attending this presentation, attendees will learn of the importance of a microfabricated device that has the power to dramatically
reduce the time associated with DNA analysis of a sample taken from a
victim of sexual assault.
This presentation will impact the forensic community by demonstrating the fabrication of a micro-total analysis system for the forensic
analysis of DNA, the time associated with the extraction, separation, and
PCRamplification of such material will decrease dramatically.
Learning Objective: The goal of this project is to develop a
macro-to-micro receptacle system that will accept a cotton swab from a
rape kit and allow for collection of desorbed cellular material. The
receptacle is to be integrated with a microchip cell separation apparatus
to circumvent conventional differential extraction.
This presentation addresses the considerable backlog of rape kit
evidence awaiting DNA analysis. Analysis of both perpetrator and
victim DNA by gel electrophoresis has been the keystone technique utilized in the investigation of cases involving sexual assault and rape, and
is a well established practice. Unfortunately, the procedures involved in
a typical DNA analysis can result in hours, even days, of laboratory time
spent on a single case, particularly in the sample preparation stages. As
a result of the time constraints involved in the analysis of such cases, and
insufficient funding, a large backlog exists in many large-volume DNA
analysis laboratories.
Microfabricated devices that utilize microchannel electrophoresis
as a DNA separation technique have been shown to greatly reduce the
time needed for analysis. The speed and efficiency of such separations
are due largely to the increased area-to-volume ratio of the etchedchannels over conventional slab gels, and the fact that high voltages can
be utilized. In addition to the aforementioned advantages, these devices
allow integration of all necessary processing steps; this provides for
complete analysis, from cellular desorption to complete DNA
assessment, onto a single microchip. Extraction of DNA from separated
sperm and epithelial cells, PCR amplification of the DNA, and separation and detection of the amplified DNA are steps that are now incorporated into single microfabricated devices. There is little doubt that
such integrated devices will transform the arena of forensic DNA
analysis.
A major component of such an integrated device is the macro-tomicro swab receptacle that must interface the “macro-scale” of the
cotton swab with the “micro-scale” of a channel used to separate
epithelial cells from sperm cells. The current protocol utilized by the
FBI for the elution of cellular materials from the cotton matrix is a time
consuming step that involves significant sample handling, which
50
directly increases the chances of sample contamination, as well as
human error. It is not unusual for a cotton swab taken from a victim of
sexual assault to be incubated for hours (overnight) in order for the cells
to elute from the cotton fibers. Additionally, the extraction solution utilizes a protein lysis buffer containing SDS and proteinase K, which aid
in the removal of the cellular material, but lyse the fragile epithelial cells
in the process. A cell-desorption process that greatly reduces extraction
time while leaving cells intact would be advantageous in that it might be
easily incorporated into a cell separation (CS) channel on a micro-total
analysis system ( -TAS).
The focus of the research presented here is the development of a
swab receptacle that provides an interface for a cotton swab taken from
a victim of sexual assault, with a -TAS on a microfabricated glass
device. Through the microscopic examination of a cotton swab containing a dry semen sample, it was discovered that the major component
leading to sperm adhesion was entanglement of the tails with the polysaccharide cellulose strands. Preliminary studies have shown that the
enzyme cellulase, which digests cellulose to produce glucose, greatly
reduced the time needed for cells to elute into solution. The desorption
of cellular material has been optimized in an aqueous solution at a temperature of 37° C in a borosilicate glass microcentrifuge tube. Eluted
sperm and epithelial cells from a cotton swab were counted hourly using
a hemacytometer, showing an approximate 2:1 ratio of sperm cells in the
presence of the enzyme in comparison with the same sample without the
enzyme. Optimum cellular elution conditions using cellulase, showing
that sperm cell desorption surpasses the current method utilized by the
FBI, as well as specifics on the integration of the receptacle with a
-TAS device will be presented.
Microchip Technology, Macro-to-Micro Interface,
Cellular Desorption
B54
Transporting or Tracking Ignitable
Liquids in Fire Scenes
Andrew Armstrong, PhD, Armstrong Forensic Laboratory, Inc., 330
Loch’n Green Trail, Arlington, TX 76012; Vytenis Babrauskas, PhD,
Fire Science and Technology, Inc., Issaquah, WA; Douglas Holmes,
MA and Cory Martin, BS*, INTROSPECT, 1023 3rd Street, League
City, TX 77573; Ray Powell, BS, The Public Agency Training Council,
Indianapolis, IN; Steve Riggs, The Public Agency Training Council,
Indianapolis, IN
The goal of this presentation is to evaluate the extent to which
transporting or “tracking” of identifiable ignitable liquids into fire
scenes occurs and offer the results of the evaluation for those conducting
scientific evaluations of fire incidents.
In the investigation of fire scenes, it is often important to determine
if ignitable liquids or liquid accelerants, e.g., gasoline, have been used.
The procedure involves collecting of debris samples and submitting
them to an analytical laboratory. If positive results are found, it may
later be necessary for the investigator to defend in court the collection
procedure used. It is occasionally claimed by individuals accused of
incendiarism that the gasoline residues found by the laboratory were not,
in fact, present initially at the fire scene but comprise an artifact.
Specifically, it has been claimed that fire investigation personnel may
have entered the fire scene having first walked through some ground
surface which was contaminated with gasoline, for example, a driveway
on which a gasoline spill had taken place. This type of claim has never
been subjected to scientific examination, thus the purpose of this work
was to create spills, have fire investigation personnel walk through these
spills, then take samples at various locations to determine if sufficient
residues can be found to obtain positive laboratory results. The locations
sampled corresponded to the footprints of the investigator directly upon
leaving the spill area. Furthermore, claims have also been made that
canines used for fire accelerant detection may also either themselves
* Presenting Author
track in sufficient contamination to cause false-positive response, or to
incorrectly alert on material that was tracked in and not originally
present. Thus, trained canines were also used in the same series of tests.
Tests have been performed to determine whether the boots or shoes
of individuals on the fire scene can track sufficient contaminants into a
fire scene to obtain a false-positive laboratory result for presence of
gasoline at the fire. Questions about the validity of forensic laboratory
results have sometimes been raised on the basis that gasoline residues
found in the laboratory samples could have been due to transport by
footwear contaminated from walking over contaminated areas. The laboratory results establish that this will not lead to false positives being
reported. Canines trained for detection of trace ignitable liquid residue
were also utilized in the test work. The results indicate that properly
trained canines show a higher sensitivity than do standard laboratory
techniques for fire debris analysis.
The results obtained in this study disprove that claim that false-positive results are likely to obtained from laboratory testing of fire-debris
samples due to contamination from the footwear of personnel. In the
present study, large quantities of fuel (and in some cases, exceedingly
large) were poured into the stepping area through which the investigator
walked prior to entering the test area. In two of the four tests, furthermore, the fuel was “raw,” that is, it was neither burned nor given
time to evaporate. Yet laboratory results were uniformly negative, apart
from samples taken from the pour location itself. The laboratory correctly identified the Test 1, 3, and 5 samples as “raw” and Test 2 and 4
samples as “weathered” gasoline. Consequently, it is concluded that
even significant carelessness by a fire investigator in entering the fire
scene with contaminated shoes will not track in sufficient gasoline that
false-positive results would be obtained.
D5S818, D7S820, D8S1179, D13S317, D16S539 and Penta D). The
Pyrosequencing technology is based on an indirect luminescent
detection of organic pyrophosphate (PPi) released from each dNTP upon
DNA chain elongation. The four nucleotides are added stepwise and following base incorporation, PPi is released and used as a substrate for
ATP sulfurase, which results in formation of ATP. The enzyme
Luciferase uses ATP to convert luciferin to its oxi-derative, which
creates a light output detectable by a CCD camera. Prior to each cyclic
addition of nucleotides, the excess of dNTPs is degraded by apyrase. The
results are shown in a pyrogram where each peak height is proportional
to the number of incorporated nucleotides.
This new technology provides a faster turnaround time of the
analysis and is highly robust. The post-PCR handling takes about 2
hours and represents a flexible platform for different types of analysis
such as; mutation detection, sequencing of short fragments, SNP and
STR analysis. The pyrosequencing method also generates a more easily
interpreted and reliable scoring of simple and complex repeats due to the
higher resolution obtained by actual sequence analysis of the repeat and
absence of common gel artefacts like stutter bands. As the peak heights
are proportional to the incorporated nucleotides, the decrease in signal
intensity by half and the specific pattern that arises after the end of the
repeat makes it possible to resolve different alleles of a heterozygous
genotype.
In this study we describe the first system for analysis of STRs using
Pyrosequencing. Eleven STR loci were analyzed among 100 Swedish
individuals to generate allele frequencies for the Swedish population.
The system has also been tested for analysis on several forensic materials from previously analyzed forensic case works.
DNA, STR, Pyrosequencing
Boots and Shoes, Contaminants, Fire Accelerants
B56
B55
Analysis of Nuclear STR Markers
Using Pyrosequencing Technology
Population Data on the Short
Tandem Repeat Loci Penta D
and Penta E in Taiwan
Marie L. Allen, PhD* and Anna-Maria Divne, MSc, Uppsala
University, Department of Genetics and Pathology, Rudbeck
Laboratory, Uppsala, 751 85, Sweden
Ling-Min Meng, MS*, Fong-Chi Wu, BS, Chung-Ming Tsao, MS, and
Chang-En Pu, MS, Ministry Justice Investigation Bureau, Taiwan, PO
Box 3562, Taiepi, Taiwan
This presentation will present a new rapid, reliable and robust technology for DNA identification in forensics.
DNA analyses in forensic investigations have made a great contribution to the possibility to convict a perpetrator or free an innocent
suspect. With the help of rapid developments in molecular technology
new methods are becoming faster and more sensitive to allow effective
and accurate DNA analyses to aid criminal investigations.
Forensic analysis has traditionally been performed using nuclear
STRs (short tandem repeats) due to the high number of alleles at each
locus. The genetic diversity of STRs makes these markers highly discriminating and suitable for individual identification in forensic investigations. DNA in forensic samples as well as in ancient DNA, however,
is often subjected to a harsh environment of degrading agents, which
might affect the ability to amplify longer fragments of nuclear DNA.
Therefore, we selected loci harbouring the shortest repeat units and
designed a PCR assay to amplify short fragments between 70-200 base
pairs. Another advantage is that the STRs were selected from the
STRBase web site, which represents markers that have been extensively
investigated for their usage in forensic applications with well-known
allele frequencies from different populations. The use of these markers
can thereby easily be transformed to new technology platforms
remaining their large informative property.
Although fragment analysis commonly used for nuclear DNA
analysis is well established and reliable, it also requires larger fragment
sizes that may be difficult to amplify on materials of poor quality and
limited amounts. In this study we have used Pyrosequencing to analyse
11 different STRs (CSF1PO, TH01, TPOX, D2S1338, D3S1358,
After attending this presentation, attendees will understand that it is
better to add the Penta D and Penta E test to the paternity test with low
CPI or add these two systems to confirm if mutations exist.
The STR loci Penta D and Penta E were observed as low-stutter and
highly polymorphic pentanucleotide repeat loci, they were thought to
add a lot portion of exclusion probability to the paternity disputes and
also to increase the discriminating probability for forensic identification,
the population data and forensic parameters of these two systems needed
to be to be studied.
Whole blood was obtained in EDTA vacutainer tubes by
venipuncture from 532 unrelated Chinese individuals and from routine
paternity cases in Taiwan. DNA was extracted by using BioGene-Fast™
30 Minute DNA extraction kit (Texas BioGene, Inc., TX, USA) and
quantitated by a Fluorometer (DyNA Quant 200, Hoefer Pharmacia
Biotech, San Francisco, CA, USA). PCR amplification was performed
by using 5ng genomic DNA in a 15£gl reaction volume comprising STR
buffer (Gold ST*R, Promega, Madison, WI USA) and 0.5 U Tag Gold
Polymerase (Applied Biosystems, Foster City, CA, USA), 1£gM
PowerPlex™ 16 primer. PCR cycling conditions followed the protocol
provided by the manufacturer. Alleles of each locus were determined
according to the ladder also provided by the manufacturer.
Aliquot of PCR products was processed by using ABI 310 Genetic
Analyzer (PE Applied Biosystems, USA). The raw data was analyzed by
the resident software (Data Collection software, version 1.0.2),
Genotypes were determined by comparing the length of the unknown
fragments to the allelic ladders provided by the manufacturer. A software
provided by POPGENE was used to analyze the data. In this study only
* Presenting Author
51
data for Penta D and Penta E were presented.
No significant deviation from Hardy-Weinberg equilibrium was
found in these systems. The most frequent allele types for each locus
were Penta D: 9(34.15%), Penta E:11(16.51%). The mean exclusion
power(MEP) for Penta E(81.92%) was larger than that of Penta
D(60.16%), and the DP for Penta E(98.6%) was also larger than that of
Penta D(93.7%). When processing paternity test and only the alleged
father were willing to be tested, the CPI was found not high enough
sometimes, but if typing results of Penta D and Penta E were added, the
CPI would increase accordingly, cases were presented with the increase
of CPI from about 7 to 17 times. We also used these two systems without
any modification on 26 Chimpanzee blood samples from Taipei
Zoo(Taiwan), there were 3 families according to the first generation
male in this group, the typing of these two systems was in accordance
with the pedigrees established by using some other 13 STR systems, the
alleles found in Penta D system were 3 and 4, the most frequently found
allele for Penta D was 4(96%), alleles 5, 7, 8, 10, 11 and 13 were found
in Penta E, and the most frequent one was 5(31%). The two Penta STR
loci described here with high MEP and PD are highly suitable for
forensic individualization and paternity tests even for the Chimpanzee.
Penta D Penta E, Population Study, Paternity
B57
Population Data on the X Chromosome
Short Tandem Repeat Loci AR, DXS10011,
DXS101, DXS6789, DXS7132, DXS8377,
DXS9895, and HPRTB in Taiwan
Meng-Yi Chen, MS* and Chang-En Pu, MS, Ministry Justice
Investigation Bureau, PO Box 3562, Taipei, Hsin-Tien, 231, Taiwan
usage of X STR.
This presentation will demonstrate the application of X STR on
special paternity cases or forensic cases.
The major applicable commercial kits for STR typing in forensic
testing are only for genomic STRs and Y chromosome linked STRs, the
typing for X chromosome linked STRs has not well developed yet, but
the forensic DNA scientists are beginning to recognize that X STRs are
powerful auxiliary systems to genomic STR, they are helpful for the
identification of female such as for differentiating if two women had the
same father directly, avoiding some of the ambiguity generated from
sibship calculation. This report contains the results of population studies
on the X chromosome STR AR, DXS10011, DXS101, DXS6789,
DXS7132, DXS8377, DXS9895 and HPRTB for Chinese living in
Taiwan. The numbers of unrelated individuals were 416 for AR, 273 for
DXS10011, 414 for DXS7132, 413 for DXS9895, 448 for DXS101, 447
for DXS6789, 450 for DXS8377 and 428 for HPRTB. The common
alleles of each locus were sequenced and used in a control ladder for
typing population samples. The primer sequences and annealing temperature were modified and optimized for designing two multiplex amplification reactions to obtain typing of all the 8 loci. For each locus 6 to 28
alleles were noted.
Heterozygosity in females ranged from 0.700 to 0.953. The Chance
of Exclusion(CE) for these 8 loci were 0.835, 0.905, 0.594, 0.562, 0.469,
0.761, 0.449 and 0.531 respectively. Among the 140 father-daughter or
mother-child pairs examined, 1 case of mutation were found at AR locus,
among the 172 father-daughter or mother-child pairs examined, 1 case of
mutation were found at DX10011, among the 216 father-daughter or
mother-child pairs examined, 1 case of mutation were found at
DXS7132.
In a deficiency case, two women claimed that they were from the
same father, the sibship index for 15 genomic STR was 1,443, and both
of the two girls had the same X STR typing AR=23/25,
DXS10011=24/30, DXS101=23/24, DXS7132=13/15, DXS8377=48/51
52
and DXS=14/15, further confirmed that they were from the same father,
in another immigration case, a grand-daughter had to be confirmed was
blood related to a grandmother(father side) to get the citizenship of
Taiwan. Because the X chromosome linked STRs would pass from
grandmother to the granddaughter, so these 8 systems were used on that,
after matching the typing of X STRs and calculating the index of
genomic STRs, their blood relationship was confirmed, this was the first
case that X STRs were used on immigration samples. These STR polymorphisms will be useful markers for parentage testing especially when
disputed child is female.
Short Tandem Repeats, X Chromosome, Population Study
B58
Validation of the Gold-Plated Silver
Sample Block on the GeneAmp®
PCR System 9700 Thermal Cycler
Melody A. Moore, MS*, John V. Planz, PhD, and A. Eisenberg,
University of North Texas Health Science Center, DNA Identity
Laboratory, 3500 Camp Bowie Boulevard, Fort Worth, TX 76107;
Rhonda K. Roby, MPH, Applied Biosystems, 850 Lincoln Centre Drive,
Foster City, CA 94404
After attending this presentation, attendees will understand that the
use of the gold-plated silver sample block is validated for use interchangeably with the silver sample block on the GeneAmp(R) PCR
System 9700 thermal cycler, and can be used for forensic casework.
This presentation will demonstrate the validation of new equipment
which is required by DAB standards. The gold-plated silver sample
block used with the GeneAmp(R) PCR System 9700 thermal cycler has
now been validated for use in forensic casework, and can be used interchangeably with other available sample block formats.
The GeneAmp® PCR System 9700 Thermal Cycler has been introduced with interchangeable silver and gold-plated silver sample blocks.
To validate the new gold-plated silver sample block on the System 9700,
amplifications were performed on both the gold-plated silver and the
previously validated silver sample blocks. PCR amplifications using the
AmpFISTR® Profiler Plus ID™, COfiler®™, Identifiler®™ and SGM
Plus®™ typing kits (Applied Biosystems, Foster City, CA) were performed. Electrophoretic characteristics such as allelic profiles, peak
heights, and peak height ratios were used to discern differences in the
amplification capabilities of the two sample blocks. There was 100%
concordance in the genetic profiles generated on each sample block.
Peak height and peak height ratio data demonstrated no differences
between the two sample blocks. These results demonstrate that the PCR
reactions on both silver and gold-plated silver blocks are equivalent. The
results of this study validate the interchangeability of the silver and goldplated silver sample blocks on the GeneAmp® PCR System 9700
Thermal Cycler.
Validation, STR, PCR
B59
Evaluation of Applied Biosystems’
RT-PCR Quantification Assays
Dixie L. Hybki, BS, MS* and John V. Planz, PhD, UNT Health Science
Center, 3500 Camp Bowie Boulevard, Fort Worth, TX 76107
After attending this presentation, attendees will understand an
improved method for quantitating DNA extracts.
This project presents a vastly improved method of DNA quantification. Additionally, one of the assays is specific for human male DNA.
This provides the ability to detect mixtures of male and female DNA
which can be beneficial in identifying samples that will be difficult to
analyze. Using these assays allows for subsequent analysis decisions
(STRs verus mitochondrial DNA sequencing).
* Presenting Author
Molecular techniques that utilize DNA require an accurate measurement of the quantity of extracted DNA. Currently, quantitation
methods include spectrophotometry, fluorometry, and hybridization
assays. Not only can some of these methods be subjective, but several
lack the ability to differentiate human versus non-human DNA templates. In a forensic setting, the DNA Advisory Board requires that a
laboratory have and follow a procedure for evaluating the quantity of
human DNA in an extract. Applied Biosystems has developed human
specific and Y-chromosome specific quantification assays using RealTime Polymerase Chain Reaction (RT-PCR). We hypothesized that
human DNA could be objectively quantitated from various tissue
sources for use in forensic work. We also hypothesized that in case of
mixtures, the male component could be quantitated independently from
the female fraction.
Comparisons were made between the total human specific RT-PCR
quantification assay and three other quantification methods: spectrophotometry, PicoGreen® fluorescent dye, and QuantiBlot™ using genomic
DNA extracted with a variety of methods and from several biological
sources. Additionally, experiments were conducted to quantify low copy
number samples using the human specific RT-PCR quantification assay.
To determine the sensitivity of the Y-chromosome specific quantification
assay, female and male DNA extracts were mixed in ratios ranging from
50:50 to 95.5:0.5 (female:male). This was followed by quantification of
DNA extracted from female epithelial fractions of vaginal swabs and of
fetal material with both the human specific and the Y-chromosome
specific RT-PCR quantification assays to determine if male DNA carryover could be detected.
Results show that DNA quantitations using ABI’s human specific
and Y-chromosome specific RT-PCR quantification assays are more sensitive and specific than current methods. Additionally, low levels of
male DNA can be detected in suspected mixture samples. We conclude
that this assay will prove valuable in quantitating human DNA and male
DNA for forensic work. This approach could prevent repeating downstream applications due to excess or minimal DNA inputs and also
provide an estimate of the extent of mixed samples.
Quantification, RT-PCR , Mixtures
B60
XTC Characterisation Using ICPMS
Gerard J.Q. van der Peijl, PhD*, Claudia P.H. van den Boom, Ing,
Annabel Bolck, PhD, and Andrew M. Dobney, PhD, Netherlands
Forensic Institute of the Netherlands Ministry of Justice, PO Box 3110,
Rijswijk, 2280 GC, Netherlands
After attending this presentation, attendees will the participant will
appreciate the possibilities and limitations of ICPMS for XTC comparison investigations.
This presentation will demonstrate new interesting forensic applications of ICPMS elemental and isotopic techniques which are presently
developed and promise to result in much more strongly discriminating
methods for forensic applications. The subject in the present presentation is just one of these applications but is very useful in demonstrating
the relevance of these techniques. The relevance is also recognized by
the international NITECRIME forensic network that has become active
in this field. More information on this network can be submitted if
requested.
Introduction. XTC (3,4-methylenedioxymethamphetamine or
MDMA) is presently one of the most favored illegal party drugs.
Important illegal production facilities are situated in the Netherlands. At
the NFI, new analytical chemical methods are being developed to characterize and compare XTC materials. Results will be presented on the
development and application of an ICPMS (Inductively Coupled Plasma
Mass Spectrometry) method for this purpose. It is expected that such
methods can be used to assist in discrimination between XTC materials
* Presenting Author
from different producers and perhaps even between different production
batches. Results from this technique will be combined with results from
other techniques such as GC-MS and XRF to maximize discrimination.
Project goal. The goal of this project was to develop a validated
forensic method and not so much to build a comprehensive collection of
XTC data. However, as part of the validation process about 100 different
XTC materials were investigated to obtain an indication of the variation
in composition and the discrimination power of the method. Almost 100
apparently unrelated XTC samples from different seizures were supplied.
Method development. Early on in the project, it was decided to
focus method development on sample digestion and ICPMS analysis of
the resulting solutions. For method development and validation purposes, a large supply of homogeneous XTC powder was prepared by
crushing and pooling a number of XTC tablets from one seizure. These
tablets were selected on the basis of availability and their anticipated
homogeneity.
Individual XTC tablets were crushed inside polythene bags, rather
than being milled, to prevent contamination from metals present in
milling equipment. The resulting powder was homogenized by coning
and quartering. 200 mg of the resulting material was placed inside precleaned quartz microwave digestion vessels into which concentrated
nitric acid (5 ml, 65% m/m) and hydrogen peroxide (1 ml, 30% v/v) were
then pippetted. These are typical ICPMS digestion reagents suitable for
many sample types and bring the elements present in the samples into
solution. The nitric acid dissolves the inorganic components whilst the
hydrogen peroxide oxidizes any organic material.
Sample digestion. Samples were digested in a closed vessel
microwave digestion unit (Multiwave, Anton Paar, Austria). The
digestion scheme was based on a published scheme used by Comment et
al. but simplified since in our system we have pressure and temperature
regulation feedback. Step 1 was a linear ramp from 700 watt to 1000 W
over 15 min., step 2 was 1000 W for 10 min. and step 3 a cooling down
period of 30 min. During step 2 the irradiation power is controlled by a
feedback loop such that a pressure of 75 bar was maintained. Under
these conditions, the temperature during step 2 was typically 280°C. The
digestion was optimized by varying the length and irradiation power of
steps 1 and 2. The concentrations of selected elements (Mg, Al, Ca, Mn,
Fe, Zn, Sr, Ba) were determined by ICP AES to arrive at the final
digestion conditions. After cooling, the samples were transferred to 50
mL plastic vessels (Sarstedt tubes) and diluted gravimetrically to volume
with Milli-Q water. Just prior to analysis, an aliquot of each sample was
diluted 10x to a final acid concentration of 3 % m/m HNO3. An internal
standard mixture (Sc, Rh, Re) was added to these final dilutions.
Literature on the inorganic composition of illicit drugs is relatively
small and even more limited for XTC. Two particularly invaluable
sources are Comment et al. [1] and Goldmann (thesis ref) [2]. From these
and other sources, the following initial list of elements, potentially
present in XTC, was derived (with isotopes to be measured): 23Na,
24,25Mg, 43,44Ca (likely present at high concentrations); 27Al, 29,30Si,
39,41K, 47Ti, 52,53Cr, 57Fe, 63,65Cu: (v possibly present at high concentrations); 7Li, 11B, 55Mn, 58,60Ni, 64,66Zn, 81Br, 85Rb, 88Sr, 96,98Mo
106,108Pd, 121,123Sb 132Cs, 137,138Ba, 194,195Pt, 197Au 202Hg, 203, 205Tl,
208Pb: (likely present at low concentrations). The anticipated concentrations are based largely on work carried out at the University of
Lausanne. The presence of these elements can be attributed to catalyst
residues, excipients, colourants etc. This list is not as extensive as for the
multi-elemental analysis of heroin or cannabis where the soil composition influences the elements found in those drugs. The synthetic nature
of XTC can be expected to preclude the kind of variation (especially rare
earth elements) found in natural drugs.
ICPMS conditions. All ICPMS experiments used a quadrupole
instrument (6100 DRC, Perkin-Elmer). Relevant instrumental parameters were: 1250 W rf forward power, 1.15 L/min nebulizer gas flow,
53
glass concentric spray chamber with a Meinhard nebulizer (sample
uptake rate ~ 1 or 0.4 mL/min depending on conditions). Daily performance and calibration experiments were performed to verify adequate
instrumental performance. The ICPMS method was developed and
refined in stages, the most important of which are described; e.g, some
of the elements in the above list were not found in any of the samples so
that a modified list was used. The influence of various other experimental parameters were investigated to result in a reliable analysis
method. The absence of matrix effects was demonstrated by standard
addition calibrations yielding the same gradients as external calibrations.
Quantification was by external calibration with internal standardization.
Results. Preliminary results of the statistical analysis of the ICPMS
data showing possible relations between the samples are presented.
References. [1] Analyse élémentaire de pilules d’ecstasy par ICP-AES et
ICPMS, Stéfane Comment, Université de Lausanne, mai 1998.
[2] These de doctorat “l’Analyse des colorants presents dans les comprimes illicites,” Till Goldmann, Licencie en Sciences forensiques de
l’Université de Lausanne, 2000.
XTC, ICPMS, ICP AES
B61
The Identification of Human Saliva
Arthur W. Young, BS* and Jennifer Regalia, MSFS, National Medical
Services, 3701 Welsh Road, Willow Grove, PA 19090
After attending this presentation, attendees will understand a rapid,
specific, and sensitive test that can be used to identify human saliva.
This presentation will a method allowing forensic scientists a single
conclusive test for human saliva rather than only presumptive tests for
saliva.
For many years, there has been no easy and conclusive test for
human saliva. In forensic practice, the test for amylase has been a commonly used method as a presumptive test for the indications of saliva.
Unfortunately, under certain circumstances, amylase can be detected in
semen, blood, and plant sources, as well as saliva from other mammalian
species, hence, the need for a more conclusive test. This study shows a
method of testing using anti-human salivary amylase that is specific to
human saliva. The test utilizes the classic antibody-antigen reaction
which results in a precipitin band. It can be run in an agarose gel using
either double immunodiffusion or crossed-over techniques. Several
species have been tested including commonly found domestic animals
such as feline and canines with no cross-reactivity. Other body fluids
and purified plant sources also show no cross-reactivity with this test.
The research in this study indicates that by using a specific anti-human
amylase antibody, a rapid, specific, and sensitive result confirming
human saliva can be obtained.
Saliva, Amylase, Antibody
B62
Internal Validation of the AmpFISTR®
Identifiler PCR Amplification Kit for
Casework Use With the ABI Prism
310 Genetic Analyzer
Steve O’Dell, MSFS*, Carl Mauterer, MSFS, Debbie Dodd, MSFS, and
Angelo Della Manna, MSFS, Alabama Department of Forensic
Sciences, 501 12th Street South, Birmingham, AL 35233
After attending this presentation, attendees will be presented with a
summary of the appropriate measures taken to implement the use of a
new capillary electrophoresis platform, the ABI Prism 310 Genetic
Analyzer, in combination with validating a new single amplification kit,
AmpFISTR® Identifiler, for the development of the 13 core CODIS
STR loci for use in forensic casework.
54
This presentation will demonstrate a complete and thorough
internal validation study according to the quality assurance standards set
forth by the Forensic DNA community for the adoption of both a new
instrument platform and multiplex kit to increase forensic DNA
casework productivity.
The ABI Prism 310 Genetic Analyzer is a single capillary automated electrophoresis platform capable of processing approximately 48
DNA samples per 24 hour period depending on the run parameters (i.e.,
sample run time, sample injection time, etc…). The AmpFISTR®
Identifiler™ PCR Amplification kit is a 5 dye short tandem repeat (STR)
multiplex assay that amplifies 15 tetranucleotide repeat loci and the
Amelogenin gender determining marker in a single PCR amplification.
Prior to casework implementation of the AmpFISTR® PCR amplification kit on the ABI Prism 310 Genetic Analyzer, internal validation
studies must be conducted. In accordance with section 8.1.2.2 (developmental validation), and sections 8.1.3.1 (a) and (b) (internal validation) of the FBI’s Quality Assurance Standards for Forensic DNA
Testing Laboratories, the Alabama Department of Forensic Sciences
completed the forensic validation of a capillary electrophoresis system
in conjunction with the AmpFISTR® Identifiler™ single amplification
kit. Results of validation studies conducted at the Alabama Department
of Forensic Sciences Birmingham DNA Laboratory showed: sensitivity
down to 100 picograms when analyzed with a 75 RFU threshold;
mixture studies exhibiting a full minor component profile at a 1:4 ratio
and a partial minor component profile at a 1:19 ratio; reproducible allele
results over 20 separate amplifications and injections conducted over a 3
day period with the 9947A positive control; and a precision of <0.13
base for all alleles. These results support the adoption of the ABI Prism
310 Genetic Analyzer for use in forensic casework with the
AmpFISTR® Identifiler™ PCR Amplification Kit at the Alabama
Department of Forensic Sciences Birmingham DNA Laboratory.
Forensic Science, Identifiler™, Internal Validation
B63
Missing Persons: Genetic Tools
That Can Help Identify Remains
Jose A. Lorente, MD, PhD*, University of Granada, Department of
Legal Medicine, Av. Madrid 11, Granada, 18012, Spain; J. Carlos
Alvarez, PhD, Carmen Entrala, PhD, Esther Martinez-Espin, MS,
Francisco Fernandez-Rosado, MS, Luis J. Martinez-Gonzalez, MS, and
Miguel Lorente, MD, University of Granada, Av. Madrid 12, Granada,
18012, Spain; University of Granada, Av. Madrid 12, Granada, 18012,
Spain; Blanca Arce, MS, Beatriz Heinrich, MS, and Jose A. Cano, MS,
Guardia Civil, Servicio de Criminalistica, DNA Laboratory, Madrid,
28003, Spain; Bruce Budowle, PhD, FBI, Laboratory Division, Senior
Scientist Biology, Quantico, VA 22135
After attending this presentation, attendees will learn the way we’re
succesfully using a DNA database to identify missing persons, and the
best ways to implement it with the maximum benefits for society and
forensic sciences.
This presentation will impact the forensic community by demonstrating the importance of non-criminal genetic databases can be of great
interest to solve social problems.
The Spanish Ministry of the Interior has implemented a National
Program to attempt to identify cadavers and bones from missing persons.
The program was named “Phoenix Program” and it began in 1999. It is
comprised of two independent genetic databases. One of the databases
is known as the Reference Database (RD). The RD initially contained
mtDNA sequences from maternally related relatives of missing persons.
The reference samples are provided voluntarily. The second database is
known as the Questioned Database (QD). The QD is comprised of
mtDNA sequences obtained from bones or cadavers that cannot be identified or that were not identified by routine and traditional procedures,
such as fingerprints, anthropology, odontology, x-rays, etc.. In all cases,
* Presenting Author
the analysis and storage of mtDNA profiles from unidentified remains
requires authorization from a judge, as mandated by Spanish law. After
mtDNA analysis provides a link (or a match) and if possible, the 13 STR
loci included in the US database (CODIS) are analyzed. Recently,
samples in the RD have begun to be analyzed for Y chromosome STR
loci (to provide paternal lineage references for analysis).
The standard operating analytical protocol is similar to that
described by Wilson et al (1995) and the nomenclature is that recognized
by the ISFG with modifications as described by Wilson et al (2002).
Once placed in the databases, DNA sequences (both nuclear and mitochondrial) automatically are compared to identify matching or related
profiles (i.e., family reconstructions), so that identifications of unknown
remains may be possible.
This process for molecular biology identification to augment other
identification procedures has been used in a number of cases in Spain,
Colombia, Chile, Peru, Mexico and the USA, with positive results. A
number of cases that could not be resolved just a few years ago by traditional anthropological means has now been solved with the use of
DNA analysis. Exemplar cases and interesting issues related to genetic
identity will be presented at the meeting.
DNA, Missing Persons, Human Rights
B64
The Missing of the Former Yugoslavia –
The Evolving Role of DNA in the
Identification Process
Edwin F. Huffine, MS*, Rijad Konjhodzic, BS, Jon M. Davoren, MS,
Adnan Rizvic, MS, and Daniel Vanek, PhD, International Commission
on Missing Persons, Alipashina 45a, Sarajevo, 71000, Bosnia and
Herzegovina
development, incorporation, and impact of a high throughput DNA
testing system on the identification efforts in the former Yugoslavia will
be detailed.
This presentation will highlight the role of DNA testing in the identification of mass disasters victims and the rapid improvement of such
technology.
During the 1990s breakup of the former Yugoslavia during, several
hundreds of thousands of people were killed, of which up to 40,000
remained unaccounted for after the end of the armed conflicts. The
International Commission on Missing Persons (ICMP) was created at
the G-7 conference in Lyon, France in 1996 with the mission of aiding
in the identification of these. As the exhumation of mass graves began
in 1996, it became apparent that the use of ‘classic’ forensic identification techniques, i.e., those not utilizing DNA testing, would frequently
be unable to establish the identity of recovered bodies. This problem
was especially pronounced in identifying the bodies from secondary
mass graves and by 1999, thousands of bodies had been recovered that
could not be identified.
The loss of life in Slovenia required only minimal DNA testing. In
Croatia, thousands of individuals lost their lives and a system was
developed in which DNA testing is an option for those cases in which
classic forensic techniques prove insufficient for identification.
However, it was for the tens of thousands of missing in Bosnia and
Herzegovina for which the evolution of the DNA process had has the
greatest impact in terms of the number of individuals identified.
Initially, no cases from Bosnia and Herzegovina were DNA tested and
the identification process was based wholly upon classic methods. Since
virtually no medical records existed in Bosnia and Herzegovina, the
accuracy of such identifications was a source of debate. Beginning in
1998, samples from a few presumptive cases were sent out of country for
DNA testing. The results obtained were used to either confirm or refute
the presumption of identity, and it was not uncommon for more than a
* Presenting Author
year to pass before DNA results were returned. The application of DNA
on such a small scale had only a minimal impact, but it did demonstrate
that DNA testing had the potential of giving answers to the families of
the missing.
In order to help address the identification process of thousands of
missing, the ICMP developed a state-of-art DNA testing system within
the former Yugoslavia, consisting of four DNA laboratories located in
Sarajevo, Tuzla, Banja Luka and Belgrade. These four DNA laboratories must work together as a system in order to bring answers to the
families of the missing. All data obtained from these four DNA laboratories is submitted to the central computer system in Tuzla, Bosnia and
Herzegovina. Additionally, eight blood collection centers were established and a comprehensive, centralized computer system was created in
which all data relating to the missing is stored. All blood and bone
samples collected in Bosnia and Herzegovina as well as the Serbia,
including Kosovo, are submitted to ICMP’s central Identification
Coordination Center (ICC) located in Tuzla, Bosnia and Herzegovina.
(It was in early 2002 when large number of bone samples began to be
submitted to the ICMP from Kosovo and later that year when bone
samples were sent to the ICMP from Serbia proper.) All samples are bar
coded at the ICC and then distributed throughout the ICMP DNA laboratory system according to the type of sample and DNA testing required.
Once a DNA report has been generated, it is given to the pathologist in
charge of the case, who is usually the person who submitted the bone
sample. It is the legal responsibility of this pathologist to contact the
family and officially close the case.
By the summer of 2003 this system was generating between 300 –
400 DNA reports per month. Once a DNA match report has been
returned to the pathologist, he/she will review ante mortem records,
articles of clothing and personal effects, and the body to ensure consistency between these ‘classic’ forms of evidence and the alleged identity
of the individual as developed by DNA testing. The magnitude and
success of this DNA testing system has altered the role of DNA testing
in the former Yugoslavia where DNA testing is now frequently used to
produce the initial lead with other identification methods assuming the
confirmation role. As a result, names are being returned to thousands of
missing.
Human Identification, ICMP, DNA
B65
Mutations in the DNA Matching
Reports of Persons Identified
Throughout the Former Yugoslavia
Ana Milos, BS*, Arijana Pozder, BS, Dijana Kadric, BS, Jon M.
Davoren, MS, Rijad Konjhodzic, BS, and Edwin F. Huffine, MS,
International Commission on Missing Persons, Alipasina 45a,
Sarajevo, 71000, Bosnia and Herzegovina
This presentation will discuss the apparent mutations that have
been observed during the course of matching the STR profiles of blood
reference samples to that of bone samples.
humanity by presenting data to describe the mutations that have been
observed during the process of identification of missing persons from
throughout the former Yugoslavia.
This presentation will discuss the apparent mutations that have
been observed during the course of matching the STR profiles of blood
reference samples to that of bone samples.
The International Commission on Missing Persons has developed a
DNA led system for the identification of the estimated 30,000 – 40,000
missing persons from throughout the former Yugoslavia. This DNA led
system involves typing of both family reference and bone samples as one
of the first steps in the identification process. Following DNA testing,
results are stored in databases and software looks for matches between
55
bone and family reference samples. This approach accelerates that identification process for samples as it uses the DNA matches as the first
piece of evidence in the process. Traditionally DNA testing has been
used to confirm identifications performed by a pathologist.
Although the identification process is accelerated by using DNA
matches as the initial lead in the identification process there are some
limitations of such a system. One limitation is the requirement to
produce reliable STR profiles from bone samples. A second limitation
is that there is a need for at least 15 STR loci in order to provide strong
statistical significance to the DNA match. A third limitation is that STR
loci can mutate and thereby reduce the statistical significance of the
match. The ICMP has been able to overcome the first limitations
through the development of extraction technique. The second limitation
has been overcome by the amplification of up to 19 different STR loci
from the Promega PowerPlex® 16 kit and the ABI SeFiler® kit. The
third limitation is much more difficult to overcome however if enough
loci are amplified they will eventually show that the suspected match is
real or just a random chance occurrence.
The ICMP has generated over 4000 DNA matching reports related
to missing persons from throughout the former Yugoslavia and is currently matching samples at a rate of 300 – 400 per month. In these
reports there have been a number for which it has been observed that the
STR profile in a parent is different from that of a child. The presence of
a single difference between related people has been observed to occur
between reference samples as well as between bone samples and the
family relatives.
STR, Mutations, PowerPlex16
B66
A Comparison of the Sensitivity of the
Power Plex®16, Identifiler®, and SeFiler®
Kits for the STR Testing of 9 – 12-Year-Old
Bone Samples Throughout the Former
Yugoslavia
Edina Omerovic, BS*, Lejla Smajlovic, BS, Jon M. Davoren, MS, Rijad
Konjhodzic, BS, and Edwin F. Huffine, MS, International Commission
on Missing Persons, Alipasina 45a, Sarajevo, 71000, Bosnia and
Herzegovina
The goal of this presentation is to compare the sensitivity levels of
the Power Plex®16, Identifiler®, and SeFiler® kits for producing STR
profiles on DNA from bone samples. These three kits have differences
in the quality of profile that is produced when used to amplify DNA from
9 – 12 year bone samples. Optimizations, of a number of parameters,
have been performed for each of these kits and will be discussed.
The breakup of the former Yugoslavia left approximately 30,000 –
40,000 missing persons. For most of these people there were no medical
records that could aid in the identification process so that left DNA
testing as the only method for identification of mortal remains. One of
the difficulties of mass testing of bone samples is that DNA isolated
from virtually every bone exhibits significant levels of degradation, contamination by microbial DNA, and inhibition of PCR amplification.
In an attempt to overcome these limitations the ICMP has optimized the Promega PowerPlex® 16, the Applied Biosystems Amplf STR
Identifiler® and the Amplf STR SeFiler® kits for the amplification of
DNA from 9-12 year old bone samples. Each of these kits has been optimized for cycling parameters, amounts of taq polymerase and magnesium concentration. Initial findings show that on average the
PowerPlex16 system amplifies the most loci however it also produces
more non specific amplification products that the other two kits.
DNA, STR, PowerPlex® 16
56
B67
Population Study on 20 Intronal SNP’s
as a Guideline for Use in Forensics
Rijad Konjhodzic, BS*, Jon M. Davoren, MS, Daniel Vanek, PhD,
International Commission on Missing Persons, Alipashina 45a,
Sarajevo, 71000, Bosnia and Herzegovina; Amina Kurtovic, BS,
University Clinical Center, Alipashina 45a, Sarajevo, 71000, Bosnia
and Herzegovina; Edwin F. Huffine, MS, International Commission on
Missing Persons, Alipashina 45a, Sarajevo, 71000, Bosnia and
Herzegovina
The goal of this presentation is to discuss the results of a SNP population study using the Applied Biosystems Taqman® assay. This study
is part of a larger project aimed at the development of a rapid and inexpensive SNP tests that provides enough discrimination to be useful as a
tool in forensic casework.
humanity by demonstrating the potential of using of SNPs in terms of
identification of victims of mass diasters.
There are currently a number of methods available for SNP testing
including Taqman®, Snapshot®, pyrosequencing, and GC-MS. Each of
these techniques has been developed for different purposes and each has
advantages and disadvantages for different uses. For forensic testing an
ideal technique would ideally be inexpensive, robust, allow high
throughput and have low requirements for DNA quantity and quality.
For some time, testing of Single Nucleotide Polymorphisms, or
SNPs, have been used as a molecular biology method, mainly for
medical research, although there have been implementations in
forensics, with various degrees of success. However, the very nature of
a SNP, as a potential single base difference, makes it attractive for
forensic scientists, who in their work are forced to deal with not only
very low quantities of DNA template, but also a degraded ones.
As a first step, the feasibility of the Taqman® assay was evaluated
as a potential forensic testing method. Twenty tested SNP’s, located in
intronal sequences and with the a lesser allele having a frequency of
above 40% were selected from the Applera SNP database. The 20 SNPs
were genotyped by 5’ nuclease reactions using TaqMan MGM probes.
The amplification, which is very simple, and allows preparation of large
numbers of samples in short periods of time. Amplifications were performed done on an Applied Biosystems 9700 master cyclers, and a post
PCR read for allelic discrimination was performed done on an Applied
Biosysytems SDS 7700.
SNPs, ICMP, DNA
B68
A Comparison of the Statistical
Significance in the Loci of the
Promega Powerplex® 16 System
From Identified Remains
Sinisa Nacic, BS*, Edina Omerovic, BS, Lejla Smajlovic, BS, Jon M.
Davoren, MS, and Edwin F. Huffine, MS, International Commission on
Missing Persons, Alipasina 45a, Sarajevo, 71000, Bosnia and
Herzegovina
After attending this presentation, attendees will get a idea of the statistical significance of each of the Promega PowerPlex® 16 loci when
used for identification of missing persons from the former Yugoslavia.
humanity by providing useful information for people using STR systems
for large scale identification work.
* Presenting Author
The ICMP has developed a DNA-led identification effort to assist
in the identification of the estimated 30,000 – 40,000 missing persons
who remain missing as a result of the breakup of the former Yugoslavia.
Due to the conditions of the recovered bodies, coupled with a relative
lack of medical and dental records, the vast majority of bodies currently
being recovered cannot be identified without the use of DNA testing.
Currently more than 10,000 bone samples and 43,000 blood samples
have been successfully tested with the Promega PowerPlex® 16 system.
The process of matching STR profiles from the reference database to
profiles in the missing person database is challenging, especially considering the large number of samples in these databases.
The vast majority of the missing has either a parent or a child as a
blood donor. Because of this fact, the ICMP has developed software that
performs an initial search based upon half-band sharing patterns. Due to
the large size of the database, random half-band sharing matches are relatively common for any given bone sample. In order to determine a
random match from a true match, additional factors are addressed. First,
blood samples are taken from multiple donors for each missing person,
and the DNA profiles from all blood donors must correspond to the
potential match with the bone sample. On average the ICMP collects
three blood references for each missing person. Once a match is found
to exist between a bone sample and multiple donors from the same
family, the statistical significance of the match is determined using
DNAview software. The DNAview software gives a likelihood ratio
(LR) for the significance of the match at each locus. Following the LR
analysis, matches that produce a posterior probability of 0.9995 or larger
are considered strong enough to generate a matching report. For the
matches where the posterior probability is less than 0.9995, additional
family reference samples are sought, which can strengthen or exclude
the match. When there are no additional family relatives available to
donate a blood sample, additional loci can be tested. It should be noted
that the DNA report does not stand alone in the identification process.
After the pathologist in charge of the case receives the DNA report, they
combine all other forensic evidence together in order to make the identification.
To examine the effectiveness of the alleles in the Promega
PowerPlex® 16 system the LR produced by each locus in the matching
reports generated by ICMP have been examined. Initial results show
that Penta E produces, on average, the strongest LR. Other loci such as
TPOX and CSF1PO produce much lower likely hood ratios in matching
reports. The substitution of the loci that produce relatively low LR ratios
for ones that produce greater discriminating potential could simplify the
identification process by reducing the number of random half-band
sharing matches that occur upon initial screening.
DNA, DNAview, Statistics
B69
The Analysis of Gamma-Hydroxybutyric
Acid (GHB) and Gamma-Butyrolactone
(GBL) in Forensic Samples Using Gas
Chromatography/Mass Spectrometry
(GC/MS) and Proton Nuclear Magnetic
Resonance (1H NMR)
Jodi E. Meyers, MS, BA*, Florida International University, 11200 SW
8th Street, CP 194, Miami, FL 33199; José R. Almirall, PhD, Florida
International University, 11200 SW 8th Street, Miami, FL 33199
After attending this presentation, attendees will be presented information on the interconversion between GHB and GBL in solution at different pH’s and time before analysis. Also, a method for the analysis of
GHB will be presented which does not cause inadvertent conversion
between GHB and GBL during sample preparation and analysis and still
maintains the sensitivity, precision and linearity of other methods currently employed.
* Presenting Author
This presentation will facilitate a more thorough understanding of
the relationship that exists between GHB and it’s lactone, GBL, in
solution. This understanding is necessary to ensure the full characterization of the sample and accurate interpretation of results. A method for
analysis of GHB is presented which does not cause inadvertent conversion between GHB and GBL and still maintains the sensitivity, precision and linearity of other methods currently employed.
Gamma-Hydroxybutyric Acid (GHB) is an endogenous compound
found in the central nervous system (CNS) and peripheral tissues. GHB,
a CNS depressant, is abused recreationally for its purported euphoric and
relaxation effects and for the purposes of drug facilitated sexual assault
(DFSA) due to its sedative and amnesic effects. The dramatic increase
in the abuse of GHB over the past decade has created the need for analytical methods to detect GHB in a variety of matrices. Specifically, the
growing use of GHB for the purposes of drug facilitated sexual assault
calls for the development of a method to determine if GHB is present in
a drink that is suspected of having been spiked.
The analysis of GHB has presented some analytical difficulties in
forensic laboratories due to the equilibrium that exists between GHB and
its lactone, GBL, in solution. Many methods currently employed may
inadvertently cause conversion between GHB and GBL during sample
preparation and analysis. A thorough understanding of the relationship
that exists between GHB and GBL under different conditions in solution
can be used to guide analytical methodology so that unintentional
sample manipulation does not occur.
In an effort to determine the effect of pH and time before analysis
on the interconversion between GHB and GBL, a study was conducted
using 1H NMR. Solutions of GHB and GBL were buffered to different
pHs (2.2, 3.1, 4.5, 6, 7.1, 8, 10) using a 1M phosphate buffer prepared in
deuterium oxide. Solutions of GHB and GBL in pure D2O were also
analyzed. The samples were stored under ambient conditions and analyzed at time zero and at selected time intervals thereafter for several
months using 1H NMR. Each solution was prepared and analyzed in
triplicate. The area of resonance lines known to originate exclusively
from GHB and GBL were ratioed to each other to determine the percentages of GHB and its lactone in solution at a particular pH and time.
pH was found to have a significant effect on the interconversion
between GHB and its lactone in solution. With this information in mind,
a method was developed that avoids sample manipulations such as pH
adjustment that could cause inadvertent conversion between GHB and
GBL. In the method developed, solid phase microextraction (SPME)
which is a fast, simple and solvent free method for the extraction of
drugs directly from aqueous samples was used for extraction and preconcentration of GHB. Extracted GHB was then derivatized on-fiber
using a silylating agent (BSTFA/TMCS). Derivatization offers several
advantages: It imparts thermal stability so that conversion of GHB to
GBL in the heated injection port of the gas chromatograph will not
occur. Also, by derivatizing GHB, a less polar and more volatile compound with better chromatographic properties is analyzed. Gas chromatography-Mass spectrometry (GC/MS) was then used for the separation and detection of derivatized GHB.
The method detects GHB in aqueous based matrices with good sensitivity, high precision, excellent linearity from 0.01 mg/mL to 0.25
mg/mL and without the need for sample manipulation that could cause
interconversion between GHB and its lactone. The method was successfully applied for detection of GHB in water as well as in several
alcoholic and non-alcoholic beverages.
Gamma-Hydroxybutyric Acid, Interconversion, Solid Phase
Microextraction
57
B70
DNA – STR Analysis of Blood Reference
Samples From Throughout the Former
Yugoslavia
Izet Eminovic, PhD* and Edwin F. Huffine, MS, International
Commission on Missing Persons, Alipasina 45a, Sarajevo, 71000,
Bosnia and Herzegovina; Nijaz Tihic, MD and Vesna Hadziavdiael,
MS, International Commission on Missing Persons, Alipasina 45a,
University Clinical Centre of Tuzla, Sarajevo, 71000, Bosnia and
Herzegovina; Jon M. Davoren, MS and Rijad Konjhodziael, BS,
International Commission on Missing Persons, Alipasina 45a,
Sarajevo, 71000, Bosnia and Herzegovina
After attending this presentation, attendees will be given an
overview of the International Commission on Missing Persons (ICMP)
high throughput blood testing facility.
humanity by presenting the lessons learned in developing a high
throughput blood STR processing lab
Attendees of this presentation will be given an overview of the
International Commission on Missing Persons (ICMP) high throughput
blood testing facility.
The International Commission on Missing Persons in the former
Yugoslavia (ICMP) has been charged with the task of identification of an
estimated 30,000 – 40,000 missing persons that remained following the
breakup of the former Yugoslavia. For almost all of the missing persons
from the former Yugoslavia the only practical approach to begin the
identification process involved DNA testing. The only feasible currently
feasible approach for this was to develop DNA STR profiles from bone
samples and match those to family reference blood samples.
To date ICMP has collected, processed, and databased more than
50,000 blood samples from family references. The DNA was extracted
from the blood samples using the S&S ISOCARD protocol. Extracted
DNA was amplified using the Promega PowerPlex 16® system and analyzed on an ABI 3100 Genetic Analyzer. Over 98% success rates were
observed for the procedure when using 50% of the recommended
amount of the Promega PowerPlex® 16 kit in 12.5 ìl reactions.
DNA, STR, Blood
B71
Purification of DNA From Sperm Cells
for Forensic Analysis Using a Sol-Gel
Filled Microfabricated Device
Joan M. Bienvenue, MS*, University of Virginia, Department of
Chemistry, McCormick Road, Charlottesville, VA 22904; Natalie
Duncalf, University of Huddersfield, Queensgate, Huddersfield, West
Yorkshire HD13DH, England; Jerome P. Ferrance, PhD and James P.
Landers, PhD, University of Virginia, Chemistry Department,
The goal of this research presentation is to demonstrate the utility
of sol-gel microchip technology for extraction and purification of DNA
from sperm cells for subsequent forensic analysis.
This presentation will describe efforts to improve conventional
extraction of DNA from sperm to both improve the efficiency of the protocol and also decrease the amount of time needed to accomplish the
assay. In addition, this work contributes to the overall goal of a fullyintegrated, microchip based DNA analysis protocol to help eliminate the
current backlog of casework.
Current techniques for DNA analysis require labor-intensive and
time-consuming processes. These methods, though effective, have led to
a dramatic backlog of casework, overwhelming crime laboratories at this
time. In addition, data-basing efforts are hindered by this backlog of
cases and, in the current condition, many cases simply go unanalyzed.
58
As such, research efforts in forensics have focused on improving the
methods associated with the analysis of DNA to develop a more rapid
and efficient assay for casework profiling.
The application of microdevices to bioanalytical analyses has drastically reduced the time required to perform a wide variety of assays. As
such, microdevices are currently being designed to improve the efficiency of processes associated with forensic casework analysis. A fullyintegrated, microchip capable of performing the steps normally carried
out on the bench-top would not only reduce the time required to perform
these tasks, but would also eliminate user intervention and potential
sources of contamination, as well as preserve more of the sample for
future analysis. PCR and high-resolution DNA separations can currently
be carried out on-chip, as well as solid-phase extraction of DNA from a
variety of clinical and biohazardous samples. In addition, current
research efforts are being directed to an on-chip differential separation
of sperm and vaginal epithelial cells, with the ultimate goal of integration of all of these processes into one, fully-functional device.
The focus of the research presented here is the extraction of DNA
from sperm cells, either directly from semen or removed from other evidentiary materials, such as vaginal swabs. Nucleic acid extractions from
biological material have historically been a laborious process, requiring
phenol-based extractions and other time-consuming methods. Recently,
a shift to solid-phase extractions on silica or ion exchange resins has not
only made DNA extractions more efficient, but these methods are also
more amenable to incorporation into microchip-based devices. We have
previously demonstrated the use of thermally-bonded borofloat glass
microchip devices packed with silica beads for the purpose of DNA
purification. Monolithic matrices prepared from a silica monomer,
tetramethoxysilane or TMOS, can be easily created in the microfabricated device format. These have been shown to yield fast, efficient,
solid phase extraction of DNA from a variety of biological materials.
The process relies on the adsorption of DNA to the solid surface via
hydrogen bonding. The work reported here investigated extraction and
purification of DNA from semen, a complex biological mixture. Also
highlighted is the extraction of DNA from a microchip purified sperm
cell fraction. A method for extraction and purification is described,
along with an elution profile for DNA from the solid phase in small
volume aliquots. A detailed evaluation of extraction efficiency of DNA
from semen using the microchip-based system is also described. The
effect of DTT, a standard component in differential extraction of sperm
cell DNA, on the extraction efficiency is presented, with conditions for
optimum extraction efficiency detailed. Comparisons of the microchip
extraction with conventional kit-based methods (Qiagen®, Promega®)
are also presented. Finally, the suitability of the DNA extracted for subsequent PCR is demonstrated by COfiler™ and Profiler Plus™ amplification, with conventional analysis on ABI 310 instrumentation. This
work represents one of the major steps required for the incorporation of
a solid phase-based extraction process for DNA into either modular or
fully integrated microdevices capable of total systematic DNA analysis
for forensic casework.
Sperm, DNA, Microchip
B72
GeneMapper™ ID Software Test Plan
Performed for Software Verification
Rhonda K. Roby, MPH*, Cherisse Boland, BS, Yasser Daoudi, BS,
Liwei Qi, MS, and Brenda Takara, MS; Applied Biosystems, 850
Lincoln Centre Drive, M/S 404-3, Foster City, CA 94404
The goal of this presentation is to summarize the verification performed for GeneMapper™ ID Software v. 3.1, an integrated fragment
analysis software application, on databasing- and forensic-simulated
samples amplified with AmpFISTR® PCR Amplification Kits.
This presentation will describe and verify an excellent tool for genotyping of samples for forensic, databasing, and parentage applications.
* Presenting Author
GeneMapper™ ID Software v. 3.1 (Applied Biosystems, Foster
City, CA, USA) was specifically designed to be a complete integrated
software solution for forensic, paternity, and databasing laboratories performing STR analysis using ABI PRISM® genetic analysis instruments
and AmpFISTR® PCR Amplification Kits for automated genotyping.
This software analyzes the raw data collected from the ABI PRISM®
instrument platforms and automatically identifies peaks, quantifies
signal intensity, sizes each DNA fragment, and makes allele calls using
defined panels and bin sets with its automated genotyping capabilities.
A test plan, which Applied Biosystems has defined as verification of the
software, was designed to develop a set of test criteria to evaluate the
performance of this software in the human identification communities.
The test plan lists approximately 50 key elements to evaluate the
software’s robustness, performance, and feature design.
Leibelt, et al. first introduced GeneMapper™ ID Software v. 3.1 at
the American Academy of Forensic Sciences Annual Meeting, 2003.
Additionally, a comprehensive concordance study of STR profiles of
forensic-simulated samples (e.g., mixtures, degraded DNA, and
inhibited DNA) generated with six (6) different AmpFISTR® PCR
Amplification kits (Identifiler®, Profiler Plus®, COfiler®, SGM Plus®,
and SEfiler™ kits), analyzed on four (4) ABI PRISM® genetic analysis
instruments, and using various ABI PRISM® software packages for both
data collection and data analysis was presented by Boland et al.
(American Academy of Forensic Sciences Annual Meeting, 2003).
Samples were processed on the ABI PRISM® 310 Genetic Analyzer (for
both Macintosh® and Windows NT® operating systems), 377 DNA
Sequencer (for both Macintosh® and Windows NT® operating
systems), and 3100 Genetic Analyzer and 3100-Avant Genetic Analyzer.
All samples were then analyzed using both GeneScan® software and
Genotyper® software and compared to GeneMapper™ ID Software v.
3.1. Data were analyzed with GeneScan® Software v. 3.7.1 and
Genotyper® Software v. 3.7, for use with Windows NT® OS;
GeneScan® Software v. 3.1.2 and Genotyper® Software v. 2.5.2, for use
with Macintosh® OS; and GeneMapper™ ID Software v. 3.1 in Classic
and Advanced modes. The combination of these studies and associated
data have led to a comprehensive verification test plan for
GeneMapper™ ID Software v. 3.1.
The verification test plan requires that features such as workflow;
data handling; peak detection and sizing; HID algorithm testing; and
CODIS functions be evaluated with a pass/fail criteria. These criteria
include: allele number error for loci containing more alleles than specified in the analysis method; out of bin alleles; peak height ratios, low
peak height and spectral pull-up levels specified by the user; broad peaks
when the width of the called peak is wider than a specified value; offscale; control concordance when the designated control sample’s
genotype does not exactly match the definition; and overlap for peaks
positioned within the overlapping size range of two markers.
GeneMapper™ ID software includes three peak detector algorithms allowing different levels of user control over data analysis. The
“classic” mode produces very similar results as those analyzed with
GeneScan® Software v. 3.1.2 designed for the Macintosh® operating
system. This algorithm aids in the adoption of GeneMapper™ ID
software for laboratories currently using GeneScan® software
developed for use with the Macintosh® OS wanting to maintain current
interpretation guidelines. The “advanced” mode provides the user with
the same analysis parameters available in GeneScan® Software v. 3.7.1,
designed for use with the Windows NT® operating system, including
several improvements made to the algorithm. An additional “basic”
mode allows for analysis using limited parameters consisting of a user
defined minimum peak height threshold. Additional features new to the
software include CODIS export functionality, automated sample concordance checking and search capability within the GeneMapper™ ID
software database. The design for verification testing and the results
from the verification testing will be presented.
* Presenting Author
ABI PRISM and its Design, AmpFISTR, Applied Biosystems,
COfiler, GeneScan, Genotyper, Identifiler, Profiler Plus, and SGM Plus
are registered trademarks of Applera Corporation or its subsidiaries in
the U.S. and certain other countries. AB (Design), Applera,
GeneMapper, and SEefiler are trademarks of Applera Corporation or its
subsidiaries in the U.S. and certain other countries. Mac and Macintosh
are registered trademarks of Apple Computer, Inc. Windows NT is a
registered trademark of the Microsoft Corporation. All other trademarks
are the sole property of their respective owners.
Software Verification, GeneMapper ™ ID Software v. 3.1,
AmpFISTR® PCR Amplification Kits
B73
It May Have Two Sides,
But It Is the Same Coin
awareness concerning the implications of law-induced asymmetries that
arise in applying science to physical evidence analyses and interpretations.
The marriage between law and science is a difficult one. Each of
these professional pursuits has a different purpose. In the physical evidence arena, lawyers may wish to control the dissemination of scientific
information generated on behalf their clients. All of the legal machinations related to access cannot be allowed to affect the objectiveness of
the science. Scientists must hold to a high standard and play no role in
manipulating the science to fit the case needs. The science must not be
compromised. The session theme of “two sides of the same coin” offers
the opportunity to contrast the perspectives of science and the law with
respect to the results of physical evidence analyses. By way of an oversimplified analogy lawyers may only see one side or the other of the
coin, or want to, while the scientist is obligated not only to recognize the
two different sides but also to see it as a single coin. Where two scientists disagree, there is a problem. Science requires objective analysis and
attempts to reach a consensus. Two competent scientists examining the
same evidence should reach the same conclusions. If this is not the case,
there is a serious problem. One or both are wrong, or the subject is not
properly the subject of a scientific inquiry. Cavalier attitudes such as
“you are entitled to your opinion and I am entitled to mine” are not
appropriate. Every effort should be made to resolve differences of
opinion. There is also a need to recognize when one is dealing with the
“opinion of a scientist” as opposed to a scientific opinion. There is a distinct difference. The former may have no scientific basis. If so, it is out
of place in any scientific report or legal proceeding.
The way this scientific information is used in a legal proceeding is
in the province of the attorneys. In criminal cases the asymmetry in the
management of information is most extreme. The prosecution as the representative of the state is obligated to make all relevant scientific information available to the defense. Evidence generated by the defense, on
the other hand may be regarded as “work product” and protected from
discovery. Details of the rules differ with jurisdiction. Experts for the
defense may have access to, and assess the results and interpretations
obtained by the state’s experts, but the reciprocity may not be observed
unless defense experts are offered as witnesses.
Science-Law Interface, Reciprocal Discovery, Criminalistics
59
B74
Evaluation of Anomalous Data
Steven J. Avato, BA*, Bureau of Alcohol, Tobacco, Firearms &
Explosives, 800 K Street, NW, Suite 710, Washington, DC 20001
The goal of this presentation is to remind forensic scientists and
criminal investigators of the importance of critically evaluating evidence
recovered from crime scenes to determine its proper significance within
the context of the overall scene. Items recovered at a scene may be
critical in determining the facts of an investigation, or may lead investigators to draw erroneous conclusions. A case study will illustrate the
serious consequences associated with improperly interpreting anomalous evidence.
humanity by serving as a reminder of the importance of critically evaluating forensic evidence in the context of the totality of a scene.
The proper evaluation of forensic evidence requires assigning a
“weight” to the importance of each piece of evidence. Investigators must
decide the significance of each item recovered from a crime scene.
Many scenes, and particularly fire scenes, present anomalous and sometimes conflicting data that must be carefully considered to determine
what role, if any, they play in establishing a scene fact pattern. The
investigation of fires, like most historical reconstructions based on
science, requires investigators to gather evidence, analyze the collected
data and form reasonable hypotheses based on the analysis of the evidence. Many times, all of the evidence converges and a single, clearly
defined origin and cause can be determined. During other investigations,
insufficient data is available on which to base a valid origin and cause
conclusion. Still other investigations present a more challenging
dilemma for fire investigators. In these cases, data is collected that may
be unexpected or unanticipated within the context of the scene. Such
findings may reasonably allow for diametrically opposed cause determinations. That two opposing conclusions can be drawn from the same
data may be anathema to forensic scientists, however Thomas S. Kuhn
stated that “Philosophers of science have repeatedly demonstrated that
more than one theoretical construction can always be placed upon a
given collection of data.”1 For example, the presence of kerosene
recovered from fire debris samples may corroborate the hypothesis that
an ignitable liquid was deliberately poured in an effort to accelerate
flame spread. However, the liquid having been spilled during the filling
of a kerosene heater might reasonably explain the same finding of
kerosene. Another hypothesis could explain the kerosene as the residue
of chemicals endemic to the scene. Additional information will be
needed in order to properly evaluate the significance of the kerosene’s
discovery. Was a kerosene heater present that might support the accidental spill hypothesis? Was the kerosene found only in baseboard wood
samples, suggesting that it might be the residue of an insecticide carrier?
Does the fire damage match that expected from the fire dynamics of an
accelerated fire? The fire investigator must thoroughly evaluate all of
the possible explanations before determining the merit of the finding
within the context of the incident scene. The key to successfully uncovering the truth in such cases lies in determining whether the unexpected
discovery is, in fact, an anomaly or a crucial piece of evidence on which
to base a new hypothesis.
Section 15.1 of the National Fire Protection Association’s (NFPA)
Guide for Fire and Explosion Investigations (NFPA 921) states that “In
some instances, a single item, such as an irrefutable article of physical
evidence … can be the basis for a conclusive determination of origin. In
most cases, however, no single item is sufficient in itself.” When evidence is observed and analyzed, investigators must make a judgment as
to the relative significance of that item. What happens to an investigation if an investigator assigns a disproportionate valuation to that
single evidentiary item? In the context of a fire origin and cause investigation, the answer to this question could be the difference between the
determination of an accidental fire cause and an intentional, incendiary
60
act. In the case study to be presented involving the investigation of a
fatal house fire, such an anomalous finding in debris samples taken from
the room of fire origin was encountered. The discovery of gasoline in
the fire debris samples was the basis for a determination that the fire was
incendiary, and consequently, the arrest of a man on capital murder
charges. A brief factual synopsis will be presented as the background for
a discussion on the weighing and evaluation of a single piece of evidence
against the totality of circumstances found at an incident scene.
1Kuhn, Thomas S. The structure of scientific revolutions – 3rd ed.
Chicago: University of Chicago Press, 1996.
Crime Scene Investigation, Fire Investigation, Critical Thinking
B75
Surviving the Stigma of a
Wrongful Conviction Case
Raymond J. Prime, PhD*, Centre of Forensic Sciences, 2nd Floor,
25 Grosvenor Street, Toronto, Ontario M7A 2G8, Canada
After attending this presentation, the participant will understand
that following a crisis over a wrongful conviction it is possible to restore
the confidence of the justice system and the capability of the forensic
laboratory.
In 1996 the Ontario Government called a Commission of Inquiry
into the wrongful murder conviction of Guy Paul Morin, in which hair
and fiber evidence played a pivotal role. As a result of that Inquiry the
Centre of Forensic Sciences shared a considerable amount of blame with
its justice system partners; the staff of the Centre, who had seen themselves as independent and impartial participants in the criminal justice
process, were particularly affected. Following the Inquiry, the Centre
became a focus of the media, the defense bar and even trial judges in
proceedings involving all of the service disciplines, not solely hair and
fiber. At the same time, though, there was a silver lining to this dark
cloud, which was the opportunity for change and improvement.
This presentation will relate how we were able to secure the support
of Government to commit resources, a chronic problem for forensic
laboratories, to implement change in management practices and accountability, quality assurance systems and reporting practices. By emphasizing our objectivity as scientists, our openness to external scrutiny with
our client groups through the creation of an advisory body comprised of
members of the whole justice community - investigators, prosecutors,
defense counsel, judiciary, and scientists - and a commitment to quality,
we have been able to restore the confidence that our organization has
traditionally enjoyed.
Wrongful Conviction, Quality, Organization Change
B76
Independent Analysis of Physical Evidence
in Criminal Cases—A Defense Perspective
Edward E. Hueske, MA*, Forensic Training & Consulting, LLC, 541
Halifax Lane, Coppell, TX 75019
After attending this presentation, attendees will understand why it
is prudent for the defense to review and/or reanalyze the physical
evidence in criminal cases.
This presentation will reinforce the need for independent examination of evidence on behalf of the defense. This is not intended to be an
indictment of law enforcement crime laboratories, law enforcement personnel or anyone else, but just serves to show the value of getting a
second opinion in any serious matter where life, death or personal
freedom are at stake and human error could be involved.
This presentation will provide the high lights of two shooting incidents in which an independent reconstruction of each of the shootings
* Presenting Author
yielded information of significance to the defense effort. These cases
illustrate that there can be aspects that support the defense theory that
can be over-looked by the original examiner. Honest mistakes and misinterpretations can also happen such that what appear to be “slam dunks”
for one particular theory may not necessarily be so.
Case 1: The first case involves a woman accused of having shot her
boyfriend as he sat across from her on a sofa-bed in a camper trailer. The
decedent had a near contact shot to the right forehead that exited behind
his right ear. The exiting bullet then passed through a partition at the end
of the sofa-bed, struck the side of the range that was behind it and fell to
the floor where the bullet was recovered.
The account of the shooting given to police by the woman had the
decedent on the floor of the camper and under a table with him shooting
himself. From this position, the bullet would have had to be traveling
upward when it struck the partition.
The crime scene investigators focused on the fact that the exit hole
in the partition was lower than the entry hole, indicating a downward trajectory. This, of course, was at odds with the defendant’s version of
events. They further pointed out that bloodstains were on the curtain
behind the sofa-bed and on top of the sofa-bed and that these too were
inconsistent with the decedent being down on the floor.
An independent examination/reconstruction was carried out in
which it was learned that the bullet actually was deflected after entering
the partition, thus giving the appearance of a downward trajectory when
the true trajectory was upward. It was also learned that the bloodstains
on the sofa-bed resulted from paramedics placing the decedent’s bloody
shirt there while attending to him.
A bloodstain on the wall down at the floor was clearly back spatter
from the gunshot. This, along with the trajectory determination, resulted
in charges against the woman being dropped.
Case 2: The second case involves a man accused of intentionally
firing a rifle directly into a crowd, striking and killing a young girl. The
girl was struck in the forehead by the bullet, a 7.62 x 39 steel post
Chinese round that did not exit her skull.
The defendant’s version of events was that he fired only warning
shots into the ground and that he never fired directly at the little girl.
Although there were a number of apparent bullet strikes in the ground at
the scene, the prosecution pointed out that a bullet hole in a window
screen behind the victim’s position indicated direct fire.
In reexamining the evidence, it was noted that the bullet that was
recovered from the victim had a flattened side. Autopsy photographs
showed a large, irregular entrance wound in the child’s forehead.
A reconstruction of the shooting indicated that a ricochet was
probable. The bullet condition, the entry wound appearance, and the
failure of the bullet to exit the victim’s head in spite of being a steel post
bullet were all consistent with a ricochet. Based upon this evidence, the
defendant was convicted of a lesser charge.
Shooting Reconstruction, Re-analysis, Physical Evidence
B77
Seeing Both Sides of the Coin
Thomas P. Shefchick, BSEE, PE*, PO Box 62284, Sunnyvale,
CA 94088
The goals of this research project are to show the difference in perspective between the prosecution/plaintiff and the defense in fire litigation.
A case study will be utilized to show the difference in perspective
between the prosecution/plaintiff and the defense for fire litigation.
Slides of the case study will be available to spark discussion. The facts,
legal issues, investigations and laboratory tests of the case study will be
* Presenting Author
given in this abstract. Members of the Academy can assume the position
of Judge, jury, litigator, insurer, investigator or laboratory analyst from
either side of the issue. Each participant should give a statement about
the role they play in the matter, what they are required by law to do and
what they did and why? Other members of the Academy or the litigators
can question the participants about the motives for their actions to gain
insight into their perspectives.
In the evening of a late summer day, a fire occurred in a commercial
property located in a summer resort area. The property was an
amusement facility for young teenagers. The facility had experienced
break-ins and vandalism prior to the fire. Insurance claims had been
made to replace damaged or missing amusement machines in the facility.
The weather had been cold and rainy all summer, which seriously
affected the income of the business. The fire was investigated by a
detective from the local prosecutor’s office. The detective determined
that the fire started in the ceiling of the building where fluorescent light
fixtures were located. He believed that a ballast of the light fixtures had
overheated and caused ignition of combustible materials in the ceiling.
This fire was a large financial loss in excess of several million dollars.
Two insurance companies insured the property and its contents. They
were on the hook to pay out several millions of dollars to the owners of
the property due to the insurance policies. Consequently, the insurance
companies sent a licensed professional electrical engineer to the fire
scene to examine the light fixtures. If the light fixtures were defective
or improperly installed, the insurance companies could subrogate against
their manufacturer or installer to get their money back. Over the years
many fires have been reported as caused by an overheated light fixture
ballast and claimants have been successful in recovering money from
ballast manufacturers and electrical contractors. A ballast is an electrical
device in a fluorescent light fixture, which raises the voltage and ignites
the gas in the light tube. It contains insulating materials, which some fire
investigators believe will flow out when it overheats and ignite combustibles beneath it. The electrical engineer examined the light fixtures
at the fire scene. He found that the fixtures and the wiring in the ceiling
were not defective. The insulating material that flowed out of the ballast
was due to the heat of the fire. The ballasts were thermally protected
units and they contained a device to de-energize them if they started to
overheat. The engineer informed the detective of his findings. The
detective told the engineer that another detective had found that the
serial numbers on amusement devices in the building did not “match”
the insurance claims. The serial numbers were for much older
equipment. Subsequently, the insurance companies sent a licensed
private investigator to inspect the fire scene because engineers are not
permitted to testify in many jurisdictions about arson. The private investigator found that the fire started on the floor beneath the burned-out
ceiling. He took samples from the floor area for laboratory analysis.
The laboratory tests indicated the presence of gasoline in the floor
samples. Consequentially, the insurance companies denied to pay the
owners fire claim. However, the owners were well respected members
of the community and had devoted in excess of 25 years of their lives to
public service. The owners retained a highly respected fire investigator
associated with a prestigious university to investigate the fire on their
behalf. He found that the fire originated in the ceiling of the building
and it was caused by wiring, which overheated. No electrical short-circuits were found but they could have been displaced by firefighting
activities. Conductors/wires coming out of the building’s main panelboard were heat damaged, its components showed evidence of electrical
arcing and its circuit breakers had not tripped opened due to the electrical overload. Samples taken from the floor were found to be negative
for gasoline by a testing laboratory. In addition, any gasoline found on
the floor area could have been inadvertently dropped there since the
facility contained gasoline powered miniature racing vehicles.
Perspective, Legal Issues, Fires
61
B78
Public Employee by Day,
Private Consultant by Night
George J. Schiro, Jr., MS*, Acadiana Criminalistics Laboratory,
5004 West Admiral Doyle Drive, New Iberia, LA 70520
After attending this presentation, attendees will learn about publicly
employed forensic scientists who are also private forensic science consultants during non-public hours.
humanity by demonstrating encouraging more actively working forensic
scientists to become private forensic science consultants during their offhours. By having a pool of these types of consultants, it will ensure that
evidence is always accurately and objectively reported and it will limit
the number of illegitimate “experts” who testify at trials.
Many publicly employed forensic scientists find themselves usually
called to court to testify on behalf on the prosecution. This is because
any exonerating or inconclusive evidence is presented prior to the trial
and the suspect’s case never goes to trial. Rarely does the publicly
employed forensic scientist testify on behalf of the defense. This is
usually reserved for retired public employees, public employees who can
work private cases, privately employed forensic scientists, university
professors, legitimate expert witnesses, and illegitimate “expert” witnesses.
This paper will examine the public employee who works private
cases during non-public work hours. It will show how and why public
employees become private consultants and it will discuss advantages
and disadvantages of this practice. It will also cover the impressions of
those who might find that their opinions are directly at odds with the testifying private consultant. Several case examples will also be discussed.
Most public employees stumble upon private casework, because, in
some cases, they are the only qualified individuals in the area with
access to the needed resources. The private casework usually begins by
contact from a private investigator who is interested in having a forensic
science examination conducted on a private or civil case. In most cases,
depending on the nature of the examination, the public employee must
get permission from their agency prior to accepting the case, especially
if it involves using public equipment or reagents. Some agencies might
allow full use of the facilities, while others might not allow the public
employee to use any equipment for personal financial gain. Some
agencies have a double standard, allowing vehicles and equipment to be
used for private security details, but not allowing the public employee to
use laboratory facilities. In some cases, public ethic boards must review
the request by the public employee to assure that no conflicts of interest
occur. In other types of cases and as the public employee’s reputation
grows, the public employee might be asked to review paperwork, photographs, and other documentation for attorneys involved in a case.
Typically, with these cases, no special resources are needed and the
examination can occur in the non-public working hours. Even with these
cases, the public employee must be sure that the private casework
doesn’t conflict with the public casework.
There are several advantages to this practice. One advantage is that
it provides a source of supplemental income. Another advantage is that
it can provide a second opinion or interpretation of the case evidence.
This ensures that all interpretations of the evidence are presented to the
court. It also provides the expert with exposure to proceedings in a
variety of jurisdictions. This will make the forensic scientist more wellrounded and might also provide him or her with new ideas and
approaches to cases. Having an expert available for the defendant at trial
also means fewer appeals if the defendant is found guilty. Another
advantage is that a publicly employed forensic scientist can also make a
fair evaluation of the evidence and analytical results based upon the prevailing forensic science standards and criteria.
There also several disadvantages to this practice. One disadvantage
is that there can be a variation in the submission of cases resulting in a
“feast or famine” case flow. Possible conflicts of interest can occur when
62
handling private cases. There is also the danger of the expert developing
a “hired gun” reputation, especially among his or her forensic science
peers. This is why the expert must always maintain objectivity when
reporting results and testifying in court. The public employee must never
compromise their public duties or it could result in the loss of public
employment. By establishing criteria and working with their public
agency, the public employee/private consultant can minimize the disadvantages and maximize the advantages.
Public employees make very good private consultants. This is
because they are experienced and they are usually current in the latest
available, practical technologies. They are also actively analyzing evidence and they are aware of the latest quality assurance standards in the
field. As a result, they cannot hold someone else to an unrealistic
standard of work. Provided the proper restraint and safeguards are taken,
a public employee can become a successful and respected private
forensic science consultant.
Public Employee, Forensic Scientist, Private Consultant
B79
Obscuring the Obverse:
The Obligations of Disclosure
Peter D. Barnett, BS*, Forensic Science Associates, 3053 Research
Drive, Richmond, CA 94806
process by which the work of any scientist is evaluated is independent
peer review. There are a number of elements that are necessary for adequate peer review, with the published report of the scientist as the
starting point for the peer review process. Various recommendations for
the content of a scientific report will be reviewed. Requirements for disclosure to enable an adequate independent review will be presented.
There is much debate among forensic scientists about their role in
exploring “the other side of the coin.” This reverse side of the coin is
important - but the obverse of the coin should not be obscured by lack of
disclosure of ultimate conclusions and underlying data.
The primary work product of a forensic scientist is the report. It is
the report that serves as the basis of most of the major decisions made
during the investigation, pre-trial litigation, and, often, trial. The decisions that are made, or should be made, based on the forensic scientist’s
findings in an investigation are made by individuals who are not technically competent and whose interests are often served by intentional or
unintentional misrepresentation of the report. It is the obligation of the
forensic scientist to issue a report that provides full disclosure of the
forensic scientist’s opinions and the underlying data and reasoning that
supports those opinions and minimizes, if not eliminates, the possibility
of misunderstanding or misrepresentation of the report.
Formats for reports are discussed in a variety of sources: Forensic
science text books, guidelines issued by various forensic science technical working groups, standardization bodies such as ASTM, and publication guidelines issued by scientific journals or professional societies.
Legal obligations as expressed in statutory requirements or case law
guidelines also attempt to define the material that must be disclosed by
litigants under different situations. The forensic scientist’s report serves
two purposes: First, the report informs interested parties of the results,
conclusions and implications of the work done by the forensic scientist.
Second, the report serves as mechanism by which another scientist can
review the work done and understand the reasons, and reasoning, behind
the opinions and conclusions expressed in the report. In the end, no one
who reads the report should be surprised by any opinions or conclusions
expressed by the scientist, either in the report or in subsequent testimony.
Another knowledgeable scientist should be able to review the report and
understand the basis for the conclusions and opinions expressed.
The obligations for disclosure are primarily based on the obligations of a scientist. Independent peer review is the process used by scientists to evaluate one another’s work, and it is the obligation of the sci* Presenting Author
entist to facilitate such peer review. The obligations of each scientist to
the process of peer review culminate in the publication of the scientific
report. Those obligations begin, however, at the earliest stages of the
scientific investigation: The collection of evidence and its preservation
for subsequent analysis; the determination of the examinations, analyses,
or experiments that are necessary; and the conduct of those operations in
a way which provides for adequate peer review are all the responsibility
of the forensic scientist.
Report, Disclosure, Peer Review
B80
Use of Principal Components Analysis in
the Individualization of Smokeless Powders
J. Graham Rankin, PhD* and Joy Cottle, BS, Forensic Science
Program, Marshall University, 1401 Forensic Science Drive,
Huntington, WV 25701; Cynthia Wallace, BS, Bureau of Alcohol,
Tobacco, Firearms & Explosives, National Research Laboratory, 6000
Ammendale Road, Ammendale, MD 20705
applicability of principal components analysis to forensic samples
This presentation will demonstrate the increased awareness of multivariate statistical analysis of large data sets in establishing similarity/dissimilarity between evidence and exemplars.
Traditionally, some smokeless powders can be identified to manufacturer and brand based on physical measurement of particle size and
shape. Chromatographic analyses by GCMS, HPLC or MEKC have
been shown to further distinguish samples. While some products are
quite similar from lot to lot, others vary widely, complicating both brand
identification and individualization to the lot level. Recent legal challenges to forensic evidence have stressed the need for statistical metrics
for similarity and/or dissimilarity for comparison of evidence and exemplars.
To meet this need, a series of over 200 smokeless powders from the
ATF smokeless powder collection have been analyzed for their
extractable organic constituents by HPLC-DAD and GCMS. Use of
multivariate statistical techniques, such as principal component analysis
(PCA), demonstrates that among double base powders, the combination
of nitroglycerin, 2,4 dinitrotoluene (2,4DNT), and dibutyl phthalate concentrations can be used to distinguish the various powders tested.
Among single base powders, the 2,4DNT, methyl and ethyl centralite
concentrations distinguish those powders. Greater than 95% of the
variance among the powders can be explained using these few analytes.
Cluster diagrams are useful in showing the degree of similarity or dissimilarity among the powders. Examples distinguishing among different
lots of the same brands will be given.
Smokeless Powder Analysis, Principal Components Analysis,
Explosives
B81
The Interpretation of Projected
Gunpowder Particle Deposition
and Impact Characteristics
Kay M. Sweeney, BS*, KMS Forensics, PO Box 8580, Kirkland,
WA 98034
After attending this presentation, attendees will understand how to
evaluate gunpowder deposit and impact characteristics so that more
complete scientifically based opinions about where the firearm was,
when it discharged on the scene, can be developed and proffered.
* Presenting Author
This presentation will help those forensic science practitioners
faced with interpreting gunpowder deposits understand that all information about the deposit must be thoughtfully developed. This data,
keeping in mind the mechanics of deposit, must be considered before
reaching a conclusion and offering an expert opinion relating to how the
deposit was produced. The more information, the more specific the conclusion can be.
One of the primary points of interest in gunpowder deposition interpretation is distance, commonly referred to as proximity testing. How
far was the firearm muzzle from the target at the time of firearm discharge?
When gunpowder particles are present on the fabric target medium,
there are four important characteristics to be considered when pondering
a conclusion specifying a distance.
1. Type of powder present, specifically, is it typical of the weapon
and ammunition suspected of being used.
2. Pattern of distribution. The powder pattern shape and how it is
oriented relative to the point of bullet penetration.
3. Density of powder particle deposition.
4. Penetration level. If powder particles have penetrated the surface
of the target medium, to what depth are they lodged?
In a recent case where a man had been fatally shot in the groin area,
his clothing was examined in the laboratory. The firearm, a Smith and
Wesson .357 magnum revolver, was found at the scene and submitted as
well. Ammunition remained in the weapon so appropriate ammunition
could be purchased and used in testing. The entry hole was in the bottom
of the zipper placket in the crotch of the victim’s black denim pants. A
heavy concentration of unburned gunpowder was deposited around the
entry hole and lesser amounts were noted out to distances of two to three
inches with individual particles further out. This pattern suggests contamination and transfer during the collection and preservation stages.
Since this was a thick area of fabric and the victim fell to, and remained
on his back, there was no blood on the exterior surface to help keep the
powder particles from migrating, however the heavy concentration at the
entry hole indicated a close range discharge. Examination of the
victim’s white cotton brief underwear revealed the presence of gunpowder at the edge of the bullet penetration hole in the crotch. The hole
in the denim pants was through an area made up of seven layers of fabric
comprising a seam and folded fabric of the bottom of the zipper placket.
Laboratory testing, using seven layers of denim fabric on one layer of
white cotton brief fabric on a firm backing, resulted in gunpowder being
deposited on the underlying white cotton when the firearm muzzle was
within 4 inches, or closer, to the target.
Under other conditions and circumstances, such on hard surface
targets, only tiny traces of gunpowder may be present making distance
determinations very difficult, if possible at all. On painted surfaces,
such as automobiles, the paint may be damaged as a result of the gunpowder particle impact. In the case of plastic surfaces, again the surface
may be damaged by gunpowder particle impact and/or exhibit heat associated with the discharge. In another recent case, an assault rifle chambered for 7.62 X 39 mm ammunition was suspected of being fired into
the upper corner of the inside surface of a door of a pickup truck. A penetrating bullet defect was found in a painted metal surface in this area.
A smoke pattern was noted on the plastic panel adjacent to the metal
surface with the bullet hole. Additionally, gunpowder particle impact
damage was noted around the bullet entry defect. Test firing into painted
metal vehicle panels was conducted until a similar damage pattern was
produced which showed that the muzzle of the firearm was within four
to six inches of the door when it discharged.
In another unrelated case, bullet damage was noted in the interior
front passenger’s side door panel of a passenger car. There was speculation that the victim of a fatal through and through shooting was shot
while sitting in this vehicle. Examination of the door panel in the laboratory revealed a gunpowder particle attached to the edge of the bullet
entry defect in an area down inside the map pocket. This evidence detail
63
ruled out the possibility of the victim being shot in a position where the
fatal bullet exited her body and penetrated the door panel.
Research should be conducted to provide data to help practitioners
properly evaluate gunpowder deposit and damage characteristics
involving, skin, bone, a variety of wood surfaces, a variety of fabrics,
leather surfaces and plastic surfaces, to name a few.
Gunpowder, Proximity, Pattern
B82
Identification of Organic Components
in Intact and Burned Black Powder
Substitutes Using GC-MS
John V. Goodpaster, PhD* and Raymond O. Keto, MFS, Bureau of
Alcohol, Tobacco, Firearms and Explosives, National Laboratory
Center, 6000 Ammendale Road, Ammendale, MD 20705-1250
After attending this presentation, attendees will learn the history of
black powder substitutes and the challenges in identifying the alternative
fuels present, particularly ascorbic acid. A newly developed method that
addresses this issue will be presented and its applicability to intact and
burned powder samples will be discussed.
This presentation will demonstrate that black powder substitutes
are likely to grow in popularity and therefore they will be more commonly encountered in improvised explosive devices. This presentation
will share a new method for analyzing these explosive samples as they
have been difficult or impossible to fully characterize in the past.
Black powder substitutes such as Pyrodex or Golden Powder were
originally designed to have improved properties relative to black
powder. For example, these propellants generate similar muzzle velocities at lower peak pressures, demonstrate lower sensitivity to friction
and shock, and generate fewer or no corrosive combustion by-products.
The latter is accomplished through reducing or eliminating sulfur
content and replacing it with other organic fuels such as sodium benzoate, dicyandiamide (DCDA), ascorbic acid, or fruit sugars. The first
product of this type was Pyrodex, which was patented in 1978 by Pawlak
and Levenson. This propellant contains KNO3 and KClO4 as oxidizers
and charcoal, sulfur, sodium benzoate, and DCDA as fuels. Since that
time, a number of commercial products have been developed that
contain ascorbic acid as an alternative to sulfur. The original patent for
Golden Powder (containing KNO3 and ascorbic acid) was granted in
1985. The Golden Powder formulation was later modified and produced
as Black Mag Powder (containing KNO3, KClO4, and ascorbic acid) by
the Arco Powder Company from early 1996 through January 1997. This
product has been subsequently re-released as “Black Mag ‘3”. Legend
Products manufactured Black Canyon Powder (containing KNO3 and
ascorbic acid) in 1996 and 1997. Clean Shot Powder (containing KNO3,
KClO4, and ascorbic acid) was introduced in 1999 and is manufactured
by Clean Shot Technologies. Most recently, GOEX briefly produced
Clear Shot Powder (containing KNO3 and cooked sugars, rather than
ascorbic acid) in 2001.
Among organic additives, ascorbic acid demonstrates the most significant chemical instability, particularly when in the presence of
moisture or metal ions. In this mechanism, ascorbic acid (aa) reversibly
degrades to dehydroascorbic acid, then irreversibly to diketogulonic
acid. Further degradation yields a vast array of compounds, including
dihydroxybutanedioic acid, 2,3,4-trihydroxybutanoic acid, glycoaldehyde and glyceraldehyde. As a result, ascorbic acid can be gradually lost from powders, making its identification difficult or even
impossible by traditional methods such as X-ray diffraction analysis or
infrared spectroscopy. As a result, various instrumental methods have
64
been developed to determine these species in clinical and food samples.
In particular, the use of trimethylsilyl (TMS) derivatization agents has
been successful in the separation and analysis of AA and its degradation
products by gas chromatography-mass spectrometry (GC-MS). In
addition, ascorbic acid and benzoate have been successfully identified in
intact Clean Shot Powder and Pyrodex, respectively, using electrospray
ionization mass spectrometry.
In this study, the TMS derivatization method was used to identify
organic fuels and their degradation products by GC-MS. Black powder
substitutes were extracted with bis(trimethylsilyl)acetamide (BSA) in
acetonitrile, which converts carboxylic acid and/or alcohol functional
groups into trimethylsilyl esters and ethers, respectively. Subsequent
analysis by GC-MS allowed for the identification of trace amounts of
ascorbic acid, benzoate, DCDA, sulfur (if present), and degradation
products such as hydroxylated carboxylic acids, furanones, and lactones.
Unburned samples of black powder substitutes such as Clean Shot,
Pyrodex, Triple Seven, Clear Shot, Black Canyon, Black Mag ‘3, and
Golden Powder were successfully analyzed with sample amounts
ranging from 2 – 20 mg. Ascorbic acid and/or its degradation products
were detected in Clean Shot, Black Canyon, Black Mag ‘3, and Golden
Powder. Of these samples, Black Canyon showed the greatest amount
of ascorbic acid degradation. In contrast, a lack of ascorbic acid but
detectable amounts of simple fruit sugars allowed Clear Shot to be differentiated from the other products. The presence of benzoate, sulfur,
and DCDA in Pyrodex and Triple Seven was also evident using this
method. Lastly, burned samples of pure ascorbic acid and various black
powder substitutes were analyzed for residual organic fuels and their
breakdown products.
Explosives, GC/MS, Ascorbic Acid
B83
A Comparison of GSR Collected
With Swabs and With Swabs
Analyzed by Inductively Coupled
Plasma Mass Spectrometry
Elzbieta Bakowska, PhD*, Lawrence A. Presley, MS, MA*, Anna
Foror, BS*, and April A. Shea, BA*, National Medical Services, Inc.,
3701 Welsh Road, Willow Grove, PA 19090
After attending this presentation, attendees will understand the possibility of analyzing GSR samples by ICP-MS regardless of the collection method (swabs or stabs)
This presentation will present the opportunity of utilization of ICPMS as a confirmation technique following the analysis of stubs by
SEM/EDX or by other non-destructive technique.
Cotton swabs are currently used for the collection of GSR prior to
the quantitative analysis by either atomic absorption (AA) or Inductively
Coupled Plasma Mass Spectrometry (ICP-MS). Stubs are traditionally
used for the collection of GSR subsequently analyzed by SEM/EDX.
Each of the analytical techniques mentioned above offers unique
advantages. Also both of those analytical techniques exhibit unique limitations. Currently the method of collection of GSR dictates which of
those techniques will be utilized for the analysis of the samples. This
presentation will compare the results of analyzing GSR specimens collected with either swabs or stubs by a single analytical method: ICP-MS.
This work demonstrates the opportunity of analyzing the GSR
samples by ICP-MS regardless of the method of sample collection. It
also presents the possibility of utilizing ICP-MS as a confirmation technique following the analysis of stubs by SEM/EDX or by other nondestructive technique.
GSR, ICP-MS, SEM/EDX
* Presenting Author
B84
Survey of Trace Elemental Contributions to
the Environment: Comparison of Samples
From a Diverse Group of Occupational
Workers With Primer Residues From Firearms
Albert B. Harper, PhD, JD*, Henry C. Lee Institute of Forensic Science,
University of New Haven, 300 Orange Avenue, West Haven, CT 06516;
Jeffery Schweitzer, PhD, University of Connecticut, Department of
Physics, Storrs, CT 06269; Jacob Trombka, PhD, Goddard Space Flight
Center, Greenbelt Avenue, Greenbelt, MD 20850; Carl Selavka, PhD,
Massachusetts State Police Crime Laboratory, 59 Horsepond Road,
Sudbury, MA 06451; Gerald Zeosky, MPA, New York State Police Forensic
Investigation Center, Washington Street, Albany, NY ; Raymond M. Kimble
III, BS , National Institute of Justice, 810 NW 7th, Washington, DC
By attending, participants will understand how trace elemental distributions on skin surfaces of different occupational segments of the population compare with trace element concentrations expected on the skin of
someone that has fired a weapon.
This presentation will impact the forensic community by advancing
knowledge of the natural variation of elemental components of GSR in
persons who have not handled a firearm.
The objective of this phase of the study is to obtain a reliable estimate
of the amount of environmental lead, barium or antimony present on the
hands of individuals who have not recently handled or discharged a
firearm. The study sample consists of individual volunteers from diverse
occupational groups who as a group have differing potentials of coming
into contact with environmental lead, barium or antimony.
The occupational groups selected for this experiment include:
1) carpenters, 2) electricians, 3) firearms examiners, 4) gas station attendants, 5) brake mechanics, 6) plumbers, 7) painters, 8) roofers, 9) police
officers, 10) machinists, and 11) x-ray technicians. A group of office
workers will serve as a control cohort.
In order to be included in the sample, the participant must not have
handled or discharged a firearm within the previous 24 hours, nor must the
participant have washed his or her hands within the previous hour. After
obtaining informed consent, each participant had either the right or left
hand swabbed twice with a cotton swab containing 4 drops of a 5 percent
solution of nitric acid. The non swabbed hand was sampled with a commercially available carbon backed sticky tape affixed to a stub intended for
Scanning Electron Microscopic (SEM) examination.
The collected samples were placed in the sample containers provided
by the manufacture of the GSR collection kit, placed in an envelope and
sealed. Analysis for lead, barium and antimony is being completed by the
National Medical Service for comparison with individuals who have discharged a firearm immediately before sampling testing. Results from these
Inductively-Coupled Plasma Atomic Emission Spectroscopic/Mass
Spectrometric (ICP-AES/MS) examinations of swabs, digested using a
10% nitric acid solution with incubation at 80o C for 2-hours will be provided for each of the cohorts.
Comparisons with the control group (office workers) will be used to
verify the method of collection and understanding of conventional environmental background. Comparison of elemental analytical results for
workers from “non-weapon” occupations will be used to determine the discrimination potential for these examinations. The results of these experiments will add to the body of knowledge necessary to properly interpret
results from similar collections and analytical procedures in cases in which
handling or firing of a firearm is at question.
This NIJ-sponsored work at the Henry C. Lee Institute of Forensic
Science at the University of New Haven is being performed in collaboration with other NIJ- and NASA-collaboratively sponsored research into
the application of remote sensing technologies for improved recognition of
trace evidence in and on crime scene evidence.
Gun Shot Residue, Environmental GSR, Trace Elements
* Presenting Author
B85
The Effect of Water Immersion
on the Analysis of the Organic
Additives in Smokeless Powder
Walter F. Rowe, PhD*, Maggie Medina, BS, and Jennifer Regalia, BS,
Department of Forensic Sciences, The George Washington University,
2036 H Street NW, Washington, DC 20052
Attendees will learn about what organic additives are present in
smokeless powders, about how these additives may be analyzed by
GC/MS and about how the analysis of the organic additives may be
adversely affected by environmental conditions.
This presentation will impact the forensic community by encouraging forensic chemists to explore the effects of environmental conditions on post-blast explosive residues.
Forensic chemists frequently analyze post-blast debris from pipe
bombs for traces of smokeless powder. If smokeless powder grains are
found they may be analyzed by Fourier transform infrared (FTIR) spectrometry to demonstrate the presence of the energetic compound nitrocellulose. Organic additives in the smokeless powder grains (for
example, stabilizers such as diphenylamine and ethyl and methyl centralite) may also be analyzed by gas chromatography, gas chromatography-mass spectrometry or micellar electrokinetic capillary electrophoresis (MECE). The organic additive suites in smokeless powders
are useful for determining the brand and lot of smokeless powder from
which the explosive filling of the bomb came. Environmental conditions
may adversely affect such analyses. This research examines the effect
of exposure of smokeless powder grains to water on the quantities of
diphenylamine and methyl dinitrobenzene isomers extracted from the
grains.
Samples of eight different smokeless powders were immersed in
distilled water for up to four weeks. Four of the powders were reload
powders from IMR Powder Company (Plattsburgh, NY): SR 4759 (lot #
L11208), IMR 4895 (lot # L7927), IMR 7828 (lot # L8208) and PB (lot
# L14547). Three were reload powders from Hodgdon Powder
Company (Shawnee Mission, KS): Hodgdon HS-7, Hodgdon BL-C(2)
and Hodgdon Titewad. The remaining powder was Solo 1000 reload
powder from Accurate Arms Company, Inc. (McEwen, TN). Small (4-6
mgm) samples of each powder were carefully weighed and then
immersed in 1 mL of distilled water for two and four weeks. At the end
of the immersion periods, the water was carefully removed from each
sample vial; the smokeless powder grains were then briefly rinsed with
100 µL of methanol; and finally 100 µL of a 0.1% (w/v) methanolic
solution of octadecane were added to each sample vial. After overnight
extraction, 1-µL aliquots of the methanolic extracts were analyzed by
gas chromatography-mass spectrometry using a Varian Saturn 2000
GC/MS system equipped with a Varian CP-SIL 8 capillary column. The
helium carrier gas flow rate was 1.0 mL/min. The injector temperature
was 250oC and a 1:20 split ratio was used. The following oven temperature program was used: The start temperature was 90oC; the temperature ramp was 16oC/min; and final temperature was 230oC. The mass
spectrometer scanned the mass-to-charge ratio range from 20 to 330.
The ratios of the peak areas of diphenylamine and the energetic plasticizers 2-methyl-1,3-dinitrobenzene, 1-methyl-2,4-dinitrobenzene, 4methyl-1,2-dinitrobenzene and 2-methyl-1,3,5-trinitrobenzene to the
peak area of the octadecane internal standard were determined.
The quantities of the energetic plasticizers and diphenylamine
extracted from the smokeless powder grains generally declined as the
period of immersion in water increased. However, for some of the
smokeless powders the quantities of organic additives extracted first
declined and then increased. This may be a real effect: the smokeless
powder grains may break down, exposing more surface area to the
methanol extractant. Because the method for quantitating the organic
additives in the smokeless powder samples may have been subject to sig-
65
nificant error, the ratios of the peaks representing the methyl dinitrobenzene isomers to the diphenylamine peak were also examined. For
two of the powders (IMR 4895 and IMR 7828) the ratio of the aggregate
areas of the peaks representing the energetic plasticizers to the diphenylamine peak fell significantly over the course of the experiment (in the
case of the IMR 4895 powder by more than a factor of two). Most of the
decline occurred in the first week of the experiment. The decline in the
quantity of the methyl dinitrobenzene isomers extracted from the
powder particles after water immersion may be due to the fact that the
solubilities of the methyl dinitrobenzene isomers in water are higher
than the solubility of diphenylamine. Changes in the relative amounts of
organic additives detected in smokeless powder grains would have a
negative impact on the determination of the brand and lot of smokeless
powder used in a pipe bomb.
Explosives, GC/MS, Smokeless Powder
B86
Matrix Effects on Explosives
Recovery and Detection
Michael E. Sigman, PhD* and Alexi Gapeev, PhD, National Center for
Forensic Science, University of Central Florida, PO Box 162367,
Orlando, FL 32816; Ralph H. Ilgner, MS, Chemical and Analytical
Sciences Division, Oak Ridge National Laboratory, PO Box 2008, Oak
Ridge, TN 37831
This paper is to present to the forensic science community results
from recent studies on the effects of sample matrix and sample holding
conditions on the recovery and detection of organic explosives trace evidence.
The results of this study provide important information concerning
the proper handling and analysis of post blast debris in order to optimize
chances of recovering trace levels of organic explosives.
Whereas many organic explosives have extremely low vapor pressures, nanogram quantities of the explosives can be lost by vaporization
from some surfaces in a matter of minutes. Laboratory experiments have
provided dramatic demonstration of the vaporization of a series of
explosives deposited on Teflon surfaces at room temperature. The series
of explosives studied included nitroesters, nitramines and nitroaromatics
(i.e., nitroglycerine, 2,4,5-trinitrotoluene, RDX, tetryl, PETN…). These
results will be discussed along with studies of the effect of the sample
matrix on the recovery of explosives. The matrices from which post blast
explosives samples are collected will typically be comprised of common
building materials and commercial surfaces such as brick, wood, plastic,
glass, metal, painted surfaces, and soils. The initial approach taken in
this study is to examine relatively “simple” matrices that offer a comparison of explosives recovery from surfaces with known properties. For
example, while glass and silica or sand are similar in many respects,
glass is nonporous and silica is porous. Similar property comparisons
and their effect on explosives recovery will also be discussed.
Sample holding conditions are an important aspect of the current
study. Preliminary results indicate that trace quantities of explosives may
be lost to the walls of the sample container at room temperature, whereas
lower temperatures can reduce the loss. Results from temperature effects
and holding time on the recovery of explosives from various matrices
will be discussed.
Explosives Analysis, Post Blast Residue, Trace Evidence
66
B87
LC/MS of Explosives:
RDX Characterization
Through Impurity Profiles
Alexi Gapeev, PhD*, Jehuda Yinon, PhD, and Michael E. Sigman,
PhD, National Center for Forensic Science, University of Central
Florida, PO Box 162367, Orlando, FL 32816
The objective of this paper is to present the forensic community a
methodology for characterization of a common explosive, RDX.
This study provides the tools for a methodology to detect trace
components, such as byproducts, precursors, degradation products and
additives in RDX, thus providing an individualizing profile or a fingerprint for a particular explosive sample. Accordingly, such a profile, when
finally developed, may be of a great value in differentiating and sourcing
samples.
Type and origin of an explosive is one the most important questions
to be addressed in a bombing investigation. RDX (1,3,5-trinitro-1,3,5triazacyclohexane) is a high power explosive that is extensively used for
military and commercial applications throughout the world. Unlike TNT,
RDX does not have isomers that may serve as markers for profiling.
Furthermore, many precursors and intermediates do not seem to survive
harsh manufacturing conditions.
Liquid chromatography - mass spectrometry (LC/MS) is an
attractive technique for the analysis of trace levels of explosives because
the analyses are conducted at room temperature thus preserving thermally labile compounds. This paper reports a study for the characterization of RDX through its impurities and degradation products profile
using LC/MS. This study will give forensic scientists the tools for the
development of a method to determine the manufacturing route and to
estimate time of manufacture. In addition to HMX that is the major
impurity in RDX, seven other compounds were found in RDX samples.
They can be divided in three groups:
1. Acetylated nitramines; common byproducts in RDX manufac
turing by the Bachman method
2. 1,3-dinitro-5-nitroso-1,3,5-triazacyclohexane; an RDX
degradation product
3. Impurities not associated with a particular manufacturing method
All these compounds contribute to a profile of an individual sample
that might lead to a fingerprint or a signature of that sample. It also can
be seen as a future tool that will facilitate relation of the sample to its
probable source due to some impurities being specific to a particular
manufacturing route. Degradation products and other impurities may
accumulate as the sample is exposed to the environment and therefore
their concentration could be used as a measure of the sample age.
Explosives Analysis, RDX, LC-MS
B88
Construction and Development of a
SPME/IMS Interface for Detection
of Explosive Compounds and Taggants
Evaluated by SPME/GC/MS
Jeannette M. Perr, BS*, Kenneth G. Furton, PhD, and José R.
Almirall, PhD, Florida International University, International Forensic
Research Institute, 11200 SW 8th Street, CP 194, Miami, FL 33199
This presentation describes the construction and optimization of an
interface for introduction of a solid phase microextraction fiber (SPME)
into an ion mobility spectrometer (IMS). The detection limits of the IMS
are calculated based on calibration and SPME equilibrium curves
created using SPME/GC/MS. The following explosives: 2,4,6-trinitro-
* Presenting Author
toluene (TNT) and 2,6-dinitrotoluene (DNT) and the following taggants:
dimethyldinitrobutane (DMNB), 2-nitrotoluene (2-NT), and 4-nitrotoluene (4-NT), were used as model compounds to evaluate the
interface. The absolute detection limits for the SPME/GC/MS method
were also calculated and based on three times the signal to noise (3S/N)
from five point calibration curves. The utility of the SPME/IMS
interface was also evaluated by sampling from different sized containers
in order to decrease the concentration of the explosive compound and
simulate real world application.
This research offers a method of screening large volumes of
enclosed space for explosive compounds and taggants using a newly
developed interface between solid phase microextraction (SPME) and an
ion mobility spectrometer. This instrumental technique offers speed,
rapid presumptive screening, field use, and easy data interpretation.
A system to sample large volumes of enclosed space such as may
be found in a large room, a cargo container, or the fuselage of an airplane
for the volatile components of explosive formulations is described.
Massive screening of luggage, personnel, and cargo is difficult due to the
small amount of material that is available for detection and the time constraints placed upon screeners. Analysis strategies for these two types of
trace samples are evaluated in order to create an effective method for
extraction, separation, analysis, and interpretation that will be suitable to
the investigators needs, i.e., a simple and rapid identification of ultralow levels of material using a field portable sampling system.
Solid phase microextraction (SPME) provides improvements over
the use over other extraction methods due to its selectivity, field portable
capability, cost, ease of use, shorter extraction times, and solvent free
extractions. Ion mobility spectroscopy (IMS) affords a low cost, rapid,
and portable method for presumptive analysis of organic materials, such
as explosives. These instruments have become widely used in our
nations airports and their installation base is very large. Ion mobility
spetrometry separates ions based on their gas phase mobility in weak
electric fields. Ionization through a radioactive B emitter at atmospheric
pressure can be controlled so only certain groups of compounds are
ionized, i.e., explosives. The IMS is known to be an extremely sensitive
technique with capabilities of detecting explosives in the low picogram
range and it is very easy to use and interpret the results. The sensitivity
of the IMS can be attributed to the negligible loss of ions or neutrals to
vacuum pumps, the walls of the cells, or recombination processes. This
presentation will briefly describe the applications of the SPME/IMS
interface to simple, rapid identification utilizing a field portable sampling system.
A Varian 3400 Gas Chromatography is coupled to a Saturn 2000
Ion Trap Mass Spectrometer detector with MSn capabilities and used as
a benchmark method of the identification of the analytes of interest. A
GE Ion Track Itemiser® 2 ion mobility spectrometer was used in conjunction with an in-house designed SPME/IMS interface. The operation
and detection limits of the interface/IMS system when used with SPME
as a pre-concentration and sampling device of large enclosed volumes
are presented and discussed.
Explosive Compounds, Taggants, SPME/IMS
B89
The NITE-CRIME Network: Development
of International Protocols for the Use of
Natural Isotopes and Trace Elements in
Criminalistics and Environmental
Forensics
JoAnn Buscaglia, PhD* and Robert D. Koons, PhD, FBI Laboratory,
CTFSRU, Building 12, FBI Academy, Quantico, VA 22135
Attendees of this presentation will learn about the NITE-CRIME
Thematic Network and results of its efforts to develop protocols for the
use of natural isotopes and trace elements in the fight against international terrorism, fraud and environmental crime.
* Presenting Author
This presentation will impact the forensic community by demonstrating international protocols for elemental analysis of forensic evidence; lessons learned from and results of interlaboratory validation
studies; validation studies that will help support legal ladmissibility of
elemental analysis methods
Characterization of evidentiary materials by their trace element and
isotopic compositions can provide valuable sourcing information, which
can be used in the fight against international terrorism, fraud, and environmental crime. To support this effort, in 2000, eight organizations representing European countries joined with three international partners to
form a Network for the development of analytical protocols for the use
of natural isotopes and trace elements in criminalistics and environmental forensics (NITE-CRIME). The principal goal of the NITECRIME Network is to be a global forum for the development of analytical protocols involving trace element and isotopic analysis of materials of forensic significance. Most of the analytical protocols being
developed involve the use of laser ablation inductively coupled plasmamass spectrometry (LA-ICP-MS), but additional studies utilizing
methods such as x-ray fluorescence spectrometry (XRF) and inductively
coupled optical emission spectroscopy (ICP-OES) are also being conducted. In addition to protocol development, the NITE-CRIME
Network also has the objectives of guiding the development and production of standard reference materials, providing guidelines for establishment of international databases of compositions of various materials,
and fostering exchange of information and training of forensic scientists
through publications, technical presentations, a NITE-CRIME website,
and training workshops. Specific materials of interest include glass,
bullet lead, human hair and nails, steel, tape, marble, sugar, bovine
serum, and selected drugs of abuse. Some of the lessons learned in organizing the Network, protocol development, and the conduct of interlaboratory validation studies will be discussed. Examples from the
instrument cross-validation study and glass and bullet lead protocols will
be used to illustrate considerations concerning sample preparation procedures, the use of mass spectrometers of differing designs, and the
selection of appropriate elements and isotopes for source discrimination
purposes.
Trace Elemental Analysis, Criminalistics, Interlaboratory
Validation
B90
The Scientific Working Group
for the Analysis of Seized Drugs
(SWGDRUG): Update
Joseph P. Bono, MA* and Thomas J. Janovsky, BS*, Drug Enforcement
Administration, Office of Forensic Sciences, 2401 Jefferson Davis
Highway, Alexandria, VA 22301
After attending this presentation, the attendee will learn about the
updates to the SWGDRUG process and become familiar with the most
recent recommendations for forensic drug analysis.
This presentation will describe familiarization with the recommended standards for forensic drug analysis.
The Scientific Working Group for the Analysis of Seized Drugs
(SWGDRUG) has been in existence since 1997. The mission of
SWGDRUG is to recommend minimum standards for the forensic
examination of seized drugs and to seek their international acceptance.
The objectives of SWGDRUG are the following:
• To recommend minimum standards for forensic drug analysts’
knowledge skills and abilities
• To promote professional development of forensic drug analysts
• To provide a means of information exchange within the forensic
drug analyst community
• To promote the highest ethical standards of practitioners in all
areas of forensic drug analysis
67
• To recommend minimum standards for drug examinations and
reporting
• To establish quality assurance recommendations
• To seek the international acceptance of SWGDRUG minimum
standards
The SWGDRUG core committee is comprised of representatives
from federal, state and local law enforcement agencies in the United
States, Canada, Great Britain, Germany, Japan, Australia, the European
Network of Forensic Science Institutes (ENFSI), the United Nations
Drug Control Program (UNDCP), Africa, and South America, a forensic
science educator, the American Society of Crime Laboratory Directors
(ASCLD), ASTM, and the National Institute of Standards and
Technology (NIST). All members of the core committee have worked
together over the past six years to build a consensus on the development
of recommendations which have impacted forensic drug analysis standards internationally.
Published recommendations have been available since 2000 to
forensic scientists around the world. These recommendations have
addressed Methods of Analysis, Education and Training, and Quality
Assurance issues. All recommendations were developed with input from
the international forensic drug analyst community.
SWGDRUG has undertaken the development of a second set of
recommendations which will address the following:
• Developing ethics, competence and certification standards for
forensic drug analysts
• Validating analytical methods for seized drugs
• Sampling seized drugs for qualitative analysis
These three sets of recommendations are currently in the discussion
phase. After the forensic science community has had an opportunity to
review all recommendations and provide comments, the revised recommendations will be adopted by the SWGDRUG core committee and
published.
This presentation will focus on the status of the recommendations
currently being developed, and enhancements to the recommendations
originally published in 2000. Representatives from the SWGDRUG
Core Committee will be in attendance to answer questions and address
concerns of those in attendance at the presentation.
Criminalistics, SWGDRUG, Drug Analysis
B92
The Effect of Silencers
on Gunshot Residues
H. Bülent Üner, PhD*, Institute of Forensic Sciences, Istanbul
University, Adli Týp Enstitüsü, Cerrahpaþa, Istanbul, 34301, Turkey;
Yüksel Soysal, BS, Istanbul Police Department, Ministry of Interior,
Turkey, Beylerbeyi Polisevi, Istanbul, 34301, Turkey; Ýsmail Çakýr,
PhD, Council of Forensic Medicine, Turkey, Adli Týp Kurumu,
Cerrahpaþa, Istanbul, 34246, Turkey; Salih Cengiz, PhD, Institute of
Forensic Sciences, Istanbul University, Adli Týp Enstitüsü,
Cerrahpaþa, Istanbul, 34301, Turkey
After attending this presentation, attendees will understand the effects
of silencers on gunshot residues (GSR).
In firearm wounding, the investigation of the clothing is very
important to determine firing distance. Firing distance determination is possible by analyzing of the gunshot residues (GSR) obtained from the
victim’s clothing.
The discharging of a firearm creates residues which evolve from the
primer, lubricants, propellant charge and elements that are present in the
bullets (volatized lead from the base of the bullet), cartridge casing, bullet
jackets and gun barrel. These products of firearm discharge collectively
referred as GSR.
Several factors affect the amount of GSR that are produced as well as
the amount of residues that reach the target. These are distance, barrel
length, propellant type, caliber etc. Also, attachments (like silencers) to the
muzzle of a weapon can alter the amount of residues that are reach the
target.
In this study we investigated the effect of silencers on the GSR. The
guns used in this study were Micro UZI submachine gun (SMG), silenced
Micro UZI SMG, Mini UZI SMG, and silenced Mini UZI SMG. The
ammunition used in the experiments was 9mm Parabellum type cartridges
produced by Turkish Machinery and Chemistry Foundation (TMCF). Shots
were fired at white cotton from a range of contact, 2, 5, 10, 15, 20, 30, 40,
60, 80cm. Sodium Rhodizonate Test was applied on the white cotton targets
to detect heavy metals containing GSR.
Except from the shot contact range, GSR was not found on the targets
for silenced Mini UZI SMG. GSR was found on the target up to the distance
15cm, when experimental shots were fired with silenced Micro UZI SMG.
Silencer, Gun Shot Residues, Sodium Rhodizonate Test
B91
Comparison of Incinerated Fabrics by
Differential Scanning Calorimetry
Pookalangaranarayana Ramakrishnan, MSc*, Central Forensic
Science Laboratory, Directorate of Forensic Science, Ministry of Home
Affairs; Ramanthapur, Amberpet PO, Hyderabad, Andhra Pradesh
500013, India
After attending this presentation, attendees will be able to compare
the fabrics that come partly burnt in the forensic examination.
This presentation will describe solving certain cases of fire and
arson on clothing, or fibers or fabrics.
Fibers and fabrics are commonly encountered in the crime examination. In an interesting case received, a piece of burnt and molten
fabric piece of clothing and a control clothing were sent to the laboratory
for comparison and analysis. It was a case of dowry burning case of a
girl in India. The amount of sample was very small wherein other conventional tests could not be carried out. In this context, the differential
scanning calorimetry was utilized in identifying the type of fabric. This
paper describes the detailed experimental study of the different type of
synthetic fabrics by differential scanning calorimetry.
Incinerated, Fabrics, Differential Scanning Calorimetry
68
B93
New Strategies for the Detection
of Condom Residues
Hans Sachs and Burkhard Rolf, Institute of Legal Medicine,
Frauenlobstraße 7a, Munich, D-80337, Germany; Wolfgang Keil, MD*,
Bauweberstr. 9a, München, Bayern 81476, Germany
The detection of residues from condom surfaces is a powerful
method to provide evidence for the use of condom during sexual intercourse. Microscopic and chemical investigations are used together. Data
on new condom markers are presented.
The detection of residues from condom surfaces in vaginal or
penile swabs is important during the investigation of sexual assault
cases. If a condom was used by the rapist, no sperm cells can be found.
The methods presented in this paper can help to provide evidence that a
questioned sexual intercourse took place.
The detection of residues from condom surfaces in vaginal or
penile swabs depends largely on the manufacturing process of the
condom that was used. Many different coatings are used by the industry.
By chemical analysis, for example, various silicone substances can be
detected. By microscopy, particles like starch granules can be observed.
If the detection of residues from condom surfaces should be used
for routine casework, it is necessary to establish the surface properties of
unused condoms. From these data it can be predicted what substances
* Presenting Author
could be expected in vaginal or penile swabs. These data should be used
to create a laboratory database, since such a database might be helpful to
identify condom residues in unknown samples. Furthermore, the time
dependence of the residue detection has to be established. For this
reason, vaginal and penile swabs from voluntary couples have to be analyzed. Since the manufacturers of condoms might change their coating
protocols, the database should be made topical from time to time to
cover the needs of forensic analysis.
We analyzed 50 different condom brands that are commercially
available in Germany. Some of these condoms were produced in the US
or in Sweden. The surface of the unused condoms was swabbed and then
microscopically investigated (HE staining). As shown earlier, on most
condom surfaces, corn starch granules can be observed. Furthermore,
synthetic particles made from polyethylene could be identified. These
particles replace more and more Lycopodium spores, which are suspected to cause allergies. In contrast to corn starch granules, polyethylene particles do not show polarization during microscopy. From a
forensic point of view, these particles have the advantage that they do
not occur naturally and that they are rarely found.
For the chemical investigation, the condom surfaces were rinsed.
After alkaline extraction, various silicone oils could be identified by
GS/MS. Substances that were added for the flavor of the condom could
not be identified with this technique. These substances seem to be less
important for the specificity of the observed chromatograms. However,
we will present data on the detection of dithiocarbamat. Dithiocarbamat
is more or less solely used for the vulcanization of rubber. This substance
could become a highly specific marker for the use of condoms.
Eight voluntary couples had sexual intercourse with different
condom brands. Subsequently, vaginal swabs were taken after defined
time periods. These swabs were analyzed microscopically and chemically. The “new” condom marker polyethylene particles could be
detected up to three days after the intercourse. Using GC/MS, we could
detect silicone oils up to three days as well. The presence of nonoxinol9 in vaginal swabs was only seen in rare cases. The detection of dithiocarbamat was not successful so far.
The microscopic detection of polyethylene particles in vaginal
swabs is a new strong evidence for the use of condoms. Besides that, the
presence of starch granules is important as well. The microscopic results
together with the chemical analysis using alkaline extraction and
GC/MS can provide evidence for the use of a condom up to three days
after the intercourse.
Condom Residues, Microscopy, GC/MS
B94
The Mother of All DNA Contracts 16,000 Sexual Assault Kits and
Counting (Part II)
Marie Samples, MS*, Robert C. Shaler, PhD, and Howard J. Baum,
PhD, New York City Office of Chief Medical Examiner, Department of
Forensic Biology, 520 First Avenue, New York, NY 10016; Karen
Dooling, MS, Nassau County Medical Examiner’s Office, 2251
Hempstead Turnpike, Building R, East Meadow, NY 11554
After attending this presentation, attendees will learn about the statistical information associated with a very large casework DNA contract.
This presentation will impact the forensic community by showing
some of the types of statistics collected during the project, others in a
similar situation may be able to use some of the information to develop
their own outsourcing projects.
This presentation will describe the data generated during a large outsourcing contract. This data includes information about the analysis of
sexual assault kits, such as the percentage of sexual assaults kits that were
“positive” for semen and/or male DNA, what sample types were positive
(vaginal swabs, anal swabs, etc.), and how often mixtures were encountered. Data will also be presented about the percentage of DNA profiles
* Presenting Author
generated from the cases, the percentage of DNA profiles that were SDIS
and NDIS eligible (the number of loci callable), and reasons why some
cases failed to yield CODIS eligible DNA profiles.
Information will also be presented about the “investigations aided”
(hits) generated as a result of the DNA profiles generated at the local, state,
and national levels: case-to-case matches, convicted offender matches, and
conviction matches.
Part I of this presentation is an oral presentation which will present
experiences gained during the course of the contract, as well as some interesting outcomes of the project such as the identification of sexual assault
patterns and the exculpation of an erroneously convicted man.
DNA, Sexual Assault Kits, Outsourcing
B95
Statistical Tools for Forensic
Analysis of Toolmarks
David Baldwin, PhD, Ames Laboratory, 9 Spedding Hall, ISU, Ames,
IA 50011-3020; Max Morris, PhD and Zhigang Zhou, BS, Ames
Laboratory, 102 Snedecor Hall, ISU, Ames, IA 50011; Stan Bajic,
PhD*, Ames Laboratory, 144 Spedding Hall, Ames, IA 50011-3020
The goal of this presentation is to provide statistical methods that will
provide a means to objectively calculate a “degree of association” between
matches of similarly produced toolmarks in order to validate the proposition that particular manufacturing methods produce marks on the workproduct (or tool) that are substantially different from tool to tool.
The statistical methods developed here may be applied to test the
assumption that no two tools are manufactured with the same surface
machining marks.
This presentation will show the results of a comprehensive statistical
study of toolmark variation produced by several manufacturing methods
(filing, grinding, whetstoning, broaching, stamping, and milling) and
present a data reduction/statistical method software tool for comparison of
the toolmarks.
There have been several studies that have shown the impact of various
tool manufacturing methods on the individuality of toolmarks and striations
produced on tools. These studies are reviewed in an article by Nichols1 and
have routinely shown that similar and/or sequentially produced tools have
toolmarks that are distinctly distinguishable. The major shortcomings of
these studies is that they concentrated on only a single manufacturing
method (e.g., broaching) and they typically used a small number (less than
10) of samples. This work extends these previous studies by conducting a
more comprehensive statistical study of toolmark variation produced by the
different manufacturing methods by expanding the number of samples
compared and generating a digital database to facilitate toolmark comparison and analysis.
Algorithms have been developed to facilitate the image matching and
are intended to mimic the process employed by an examiner (i.e., rotate the
samples on a comparison microscope so that principal features in each are
oriented and adjust the “hairline” separating the split image in an attempt to
find a strip along which the images exhibit similar patterns). The algorithms focus on several key steps: i) analysis of each image to determine
directions of maximum and minimum variation at a given point; ii) comparison of one-dimensional image “strips” along the directions of greatest
spatial variation; iii) iterative strategy for selecting local areas in each
image for comparison, and; iv) match validation, i.e. determining whether
an apparent match is real or an artifact due to similarities only in sub-areas
of the images.
Results of the algorithms applied to image data sets (of no less than
100 images per manufacturing process) will be presented and discussed.
Toolmark variations in manufacturing methods will be examined on commercial tools and on in-house sequentially produced samples.
1). Nichols, R.G., “Firearm and Toolmark Identification Criteria: A Review
of the Literature,” Journal of Forensic Sciences 42, 466-474 (1997).
Toolmarks, Statistics, Digital Imaging
69
B96
Eliminating Sources of Pipetting
Error in the Forensics Laboratory
David Epstein* and Shannon Boyd, NFSTC National Forensic Science
Technology Center, 7881 114th Avenue North, Largo, FL 33773
After attending this presentation, attendees will understand the role
of training in the accurate dispensing of liquids using an air displacement
pipette.
This presentation will provide a broader understanding of the topic
that will improve the quality of analysis.
Hypothesis: Pipettes are complex precision instruments that are
subject to error due to mechanical failure and improper operator technique. Improved performance may be obtained by following essential
quality control guidelines and by providing basic pipetting technique
training.
Content: This poster discusses the nature of pipette performance
including how pipettes work and why pipettes fail. Suggestions are
offered for mitigating sources of error via straightforward quality control
and operator training guidelines. Data is presented showing the beneficial impact of pipetting technique training. Specific pipetting technique training tips are provided.
Conclusion: Today’s air displacement pipettes are precision instruments that should not be taken for granted. Pipettes tend to fail silently
and randomly, impacting sample and reagent delivery. Periodic calibration and preventive maintenance are essential to ensure the integrity
of laboratory results. Operator technique training, especially if it offers
the opportunity to obtain immediate feedback, is easy to do and has a
significant beneficial effect on performance.
Pipette, Accuracy, Training
B97
Mitochondrial Coding and D-Loop
Analysis Using Pyrosequencing
Marie L. Allen, PhD* and Hanna Andréasson, MSc, Uppsala
University, Department of Genetics and Pathology, Rudbeck
Laboratory, Uppsala, 751 85, Sweden; Uppsala University, Rudbeck
Laboratory, Uppsala, 751 85, Sweden
This presentation will describe a fast analysis tool for mtDNA
analysis including extended information retrieved by analysis of coding
regions in the mitochondrial genome.
In forensic analysis, mitochondrial DNA is often used when the evidence material contains limited amounts of nuclear DNA. We have
developed a novel, rapid and easy to use typing system for analysis of
mitochondrial DNA based on the Pyrosequencing technology. This is a
rapid, non-electrophoretic, single-tube sequencing-by-synthesis method
in which a cascade of enzymatic reactions yields detectable light. The
pyrosequencing system was first developed for analysis of the highly
polymorphic mitochondrial D-loop and a database of D-loop sequences
from 200 randomly selected blood donors has been compiled. However,
a more discriminatory mitochondrial DNA analysis would be preferable,
especially in cases where none or few differences in comparison to the
Cambridge reference sequence are found between different individuals.
Therefore, the system has been extended to also include analysis of short
fragments in the coding region.
The coding region fragments were chosen, based on the diversity
determined in 26 Swedish as well as 52 European whole mitochondrial
sequenced individuals, and cover the most informative polymorphic
sites throughout the entire mitochondrial genome. A database over
coding region variation in 150 individuals has been compiled for the 19
different coding region fragments that were designed and optimised for
pyrosequencing analysis. After the initial database analysis samples with
none or one difference from the Cambridge reference sequence in the Dloop were studied in more detail. In order to achieve a better discrimi70
nation than D-loop analysis alone between these individuals, 8 of the 19
coding sequence fragments were chosen as a part of the complete typing
system. The fully developed typing system, for an optimal discrimination, consists of 18 PCR fragments and 24 pyrosequencing reactions:
8 D-loop reactions, 8 haplogroup determining reactions and 8 reactions
analysing additional SNPs for a possibility to distinguish between individuals identical in the D-loop sequence.
The pyrosequencing D-loop typing system has been evaluated in
analysis of 50 previously sequenced forensic casework evidence materials. All pyrosequencing results were identical to the Sanger sequences
with an average read length of 45-100 nucleotides. The coding fragment
analysis has shown a capability to reduce the number of not distinguishable D-loop analysed samples (due to only one or no differences)
by as much as 80%. Moreover, coding region analysis has been used
successfully in a forensic case from 1952 to exclude a suspect exhibiting
only one D-loop difference in comparison with the evidence material. In
the future, this system, for analysis of the most informative parts of the
whole mitochondrial genome, will be more informative and faster, than
the D-loop sequence analysis used routinely today.
mtDNA, Pyrosequencing, Coding mtDNA
B98
A Micellar Electrokinetic Screening
Method for Common Sexual Assault Drugs
Sandra C. Bishop, BS, Margaret A. Lerch*, and Bruce R. McCord,
PhD, Ohio University, 136 Clippinger Laboratories, Athens, OH 45701
The objective of this analysis was to develop a simultaneous
method of detection for benzodiazepines and gamma-hydroxybutyric
acid (GHB) analogs using micellar electrokinetic chromatography.
Much attention has been given to benzodiazepines and gammahydroxybutyric acid (GHB) related compounds owing to their widespread use as date-rape drugs. The rapid metabolism and lethargic
effects of these compounds make them perfect weapons for an assailant.
Many of the current screening methods for benzodiazepines involve
immunoassays that have insufficient cross reactivity with low-dose
metabolites, while current methods for the detection of GHB can involve
complex derivatization. Since either of these drugs is possible in a case
of drug facilitated sexual assault, toxicologists would greatly benefit
from a screening method that allows the simultaneous detection of both
groups of substances.
Capillary zone electrophoresis (CZE) is often compared to gas
chromatography (GC). Advantages of using CZE over traditional GC
are that it does not require extensive extraction or derivatization steps.
While CZE is limited in its ability to detect neutral compounds, micellar
electrokinetic chromatography (MECC) permits the analysis of
uncharged molecules by providing a secondary separation through the
addition of a surfactant that forms into micelles. These aggregates will
interact with the analytes of interest and carry them countercurrent to the
electroosmotic flow, enabling greater separating power.
Optimization of the borate buffer was carried out using a set of
standards containing 8 benzodiazepines, GHB, GBL, and the internal
standard, sulfanilic acid. The optimal buffer was found to contain 20
mM SDS and 7% acetonitrile at a pH of 9.2. In the case of an interfering
peak, the buffer containing 30 mM SDS and 10% acetonitrile may
provide a better separation.
The benzodiazepines were calibrated with concentrations ranging
from 2.5mg/µL to 100µg/mL. GHB was run at 0.1 to 2.5 µg/mL, while
the GBL concentrations were from 0.5 to 10 µg/mL. The method was
shown to have a detection limit of less than 2 µg/mL for five out of eight
benzodiazepines. The detection limits for GHB and GBL were 32 µg/mL
and 150 µg/mL, respectively.
In many cases of drug-facilitated sexual assault, the drug has been
placed into a victim’s beverage. For this reason, a series of GHB spiked
beverages were monitored to determine the effects of time on the drug
* Presenting Author
analysis. After 48 hours, there was no observed degradation of GHB in
any of the analyzed beverages. Possible interfering peaks from drugs of
abuse and artifacts from a variety of different drink combinations were
also studied in detail. The analysis of GHB involved a simple 1:10
dilution of the beverage sample and so any interferences were present at
very low levels. In the beer sample, there was a peak that eluted with
the GHB. Using the 30 mM SDS/10% acetonitrile buffer, the two were
separated. For the benzodiazepines, a liquid-liquid ethyl acetate
extraction was used. Using the two optimized buffers, it was shown that
a variety of interfering drugs have distinguishable mobilities in comparison to GHB and the benzodiazepines.
The method shows good separation of all benzodiazepines as well
as GHB and provides a rapid screening for many of the common sexual
assault drugs and other club drugs. When encountering an unknown
sample, the recommended procedure is to dilute an aliquot 1:10 as
described for GHB analysis. A second aliquot should be prepared for
extraction to detect the drugs that may be present at lower concentrations. We believe this method would provide an excellent overall
screening tool for the detection of date-rape drugs.
Benzodiazepines, GHB, MECC
B99
Shampoo Investigations in Relation
to a Rape Investigation
Gerard J.Q. van der Peijl, PhD* and René P. de Bruijn, Ing,
Netherlands Forensic Institute of the Netherlands Ministry of Justice,
PO Box 3110, Rijswijk, 2280 GC, Netherlands
After attending this presentation, attendees will the participant will
appreciate the possibilities of surfactant compositions for forensic investigations of toilet products such as shampoo.
humanity by demonstrating surfactants which are used in many industrial products can be encountered as forensic evidence and needs to be
characterized and compared to other evidence material. Presently surfactant component information is hardly used at all for forensic investigations. The results of this presentation demonstrate the possibilities in
using this information. It is expected more forensic applications will
follow. By making forensic practitioners aware of these possibilities it is
expected they will access this source of information.
Introduction. Results are presented of shampoo investigations in
relation to a suspected anal rape of a young boy by an older teenage boy.
The victim stated that the suspect used shampoo as a lubricant during the
rape. Victim’s anal swabs, the victim’s clothes and underwear, the
suspect’s underpants as well as all surfactant containing products from
the suspect’s bathroom were submitted by the police. In this investigation we used visual methods, LC-MS, GC-MS and MALDI MS techniques to detect and identify surfactants as well as other relevant
chemical components.
The victim and suspect’s samples were first investigated for the
presence of potential shampoo components. Afterwards component
compositions on the various items were compared. We focused at first
on involatile surfactant components. Investigations were performed
using a Crimescope system for visual characterization at a number of
wavelength regions, GC-MS for component analysis, LC-MS and
MALDI MS for surfactant analysis. Sodium lauryl ether sulphate is a
surfactant that is widely used in shampoos. All shampoos in the submitted products contain this ingredient according to the product label.
The chemical formula CnH2n+1-(OC2H4)xOSO3Na where x denotes the
number of ethylene oxy-segments and n=12 for the pure chemical form.
This is a synthetic chemical component not produced naturally by plant
or animal and not secreted by humans. Even one industrial single surfactant consists already of a large number of similar components dif* Presenting Author
fering, e.g., in the exact alkyl group (in the above example n) and the
number of ethylene oxy segments (x in the example) in a poly ethylene
oxy segment. This variation offers possibilities in a forensic investigation to determine the presence of these component mixtures and make
comparisons. Apart from the product label information further confidential product information was obtained from a number of shampoo
producers.
Visual investigation. The victim and suspect’s clothes were investigated visually for potential shampoo residues. Samples of the products
from the bathroom of the suspect were dried and investigated similarly.
Both the tops of the anal swabs and the white cotton front inside of the
suspect’s underpants displayed fluorescent stains.
Using a Crimescope (irr. 515 nm, det. > 550 nm) a light yellow
pattern was observed at the same position on the front inside of the
suspect’s underpants. On the victim’s clothes no clear stains could be
observed in daylight. For the victim’s clothing stains were especially
expected on the (rear inside of the) underpants. Further investigations of
the victim’s clothing were therefore directed on the underpants.
Only two shampoos from the relevant toilet products (including all
shampoos) from the suspect’s bathroom displayed similar fluorescence
behavior as described above. Only one shampoo (named A below) displayed a light yellow color upon drying. Application of the two fluorescent shampoos on the green material of the victim’s underpants after
drying resulted in stains that were not visible in day light and did not fluoresce using the Crimescope.
LC-MS. Instrumental conditions were selected to detect and
identify anionic surfactants such as lauryl ether sulphate. Using these
conditions mostly groups of compounds with different alkyl terminal
groups will be separated. Compounds with identical alkyl terminal group
but with different numbers of ethylene oxy segments in the poly ethylene
oxy segments can be discriminated by mass spectrometry. In a preliminary separate Flow Injection Analysis (FIA) MS experiment, bypassing
the LC separation part, it was determined that the bathroom products not
investigated in the above fluorescence experiments did not contain
sodium lauryl ether sulphate.
Methanol extracts of different fluorescent parts of the white cotton
inside on the front of the suspect’s underpants as well as the cotton
swabs with material from the victim’s anus (anus swabs) and some
selected parts of the victim’s underpants were investigated using the LCMS method. In all these different samples lauryl ether sulphate was identified with the alkyl part varying in the range decyl to hexadecyl (n=10,
12, 14, 16) and the number of ethylene oxy units, dependant on the alkyl
group, varying in the range x=0 to x=12. The results for the clothing
samples and the victim’s anus swabs corresponded well in lauryl ether
sulphate composition. Methanol diluted samples of the two fluorescent
shampoos were investigated using this LC-MS method. Results for
shampoo A corresponded well with the results for both underpants and
the anal swabs. The alkyl distribution for the other shampoo (named B
below) was somewhat narrower.
MALDI MS. This technique was applied to water extracts of the
suspect’s underpants and an anal swab from the victim. Also samples of
the two shampoos A and B were investigated using this technique.
Experiments were performed for us using facilities at Unilever Research.
Best results were obtained detecting positive ions and using a Tetrakis
matrix. Four different ion series were detected. Two ion series are interpreted as C12H25-(OC2H4)xOSO3Na2+ and C12H25-(OC2H4)xOSO3NaK+
ion-series. These were identified in the water extracts of the suspect’s
underpants, the anal swab and both shampoos. A third, as yet unidentified
ion series (m/z= 413, 475, 519, 563, 607, 651, 695, 739 and 783) was
detected in all the above samples except shampoo B. The fourth non-specific ion series has not been interpreted.
GC-MS was applied to methanol extracts of suspect and victim’s
underpants and an anal swab of the victim. Samples of the two shampoos
A and B were diluted in dichloromethane and investigated with a
standard GC-MS method. Mainly non-specific background components
71
are detected in the clothes and anal swab samples. These are identified
as compounds introduced during sample preparation and compounds
already present in blank clothing and swab samples. Among this background extra compounds were detected in the methanol extracts of the
suspect’s and victim’s underpants and the anal swab. These extra compounds are not observed in blank samples or the sample of shampoo B
but are again observed in the sample of shampoo A. For now these compounds have not been identified.
Conclusions:
1. Lauryl ether sulphate components were detected on the
suspect’s and victim’s underpants as well as the anal swabs.
Lauryl ether sulphate is an anionic surfactant that is widely
used for shampoos and shower/bath products. The human body
does not secrete it.
2. The lauryl ether sulphate components were detected locally on
the inside front of the suspect’s underpants. The distribution of
the lauryl ether sulphate components on the victim’s underpants
was less clear, partly as a consequence of different fluorescence
product behavior on this underpant.
3. Shampoos A and B are the only products of the 13 products
from the suspect’s bathroom that correspond in some
characteristics with the residues on both suspect’s and victim’s
underpants and the anal swabs.
4. Results from visual, LC-MS, MALDI MS and GC-MS
investigations on the samples of victim and suspect correspond
better with shampoo A than with shampoo B.
In our opinion the residues on the suspect’s underpants and the
victim’s anal swabs are from the same category of products, probably a
shampoo, shower or bath product. Out of all the products in the suspect’s
bathroom shampoo A is the most likely candidate for this.
Shampoo, Surfactants, LC-MS
B100 Performance Testing of Commercial
Containers for Collection and Storage
of Fire Debris Evidence
Mary R. Williams, BS*, National Center for Forensic Science,
University of Central Florida, PO Box 162367, Orlando, FL 32816;
Denise Fernandez, University of Central Florida, PO Box 162367,
Orlando, FL 32816; Michael E. Sigman, PhD, National Center for
Forensic Science, University of Central Florida, PO Box 162367,
Orlando, FL 32816
The objective of this paper is to present to the forensic science community results from recent studies on the performance of commercial
containers in the retention of ignitable liquids.
This presentation will provide important information concerning
the proper handling and analysis of fire debris evidence and the viability
of commercial sample containers.
Long-term storage of samples in suspected arson cases requires that
the evidence containers be free from leaks that can result in total loss of
the volatiles or evaporative aging of the sample, leading to an altered
hydrocarbon profile. Previous investigations have reported significant
leak rates for various commercial containers, primarily paint cans and
polymer bags. This research project undertakes a study of the vapor
retention properties of commercial containers through repeated removal
of small (c.a. 20 micro-L) vapor samples from large (c.a. 1 gal.) containers, and an investigation into the dynamic behavior of hydrocarbon
vapors in the closed container.
The dynamic behavior of a mix of volatile hydrocarbons inside a
closed container complicates analyte sampling and analysis, while at the
same time revealing important properties of the vapor which can significantly impact the interpretation of analytical data. Depth profiling of
72
recovered hydrocarbons within closed containers by collection on
standard carbon strips will be presented for a set of hydrocarbons, and
the practical implications of the data for sampling methodology and
source determination will be discussed.
Results from a long-term study of hydrocarbon retention by one
gallon commercial paint cans will be presented. Data revealing the leak
rate of the containers and the dynamic behavior of the hydrocarbon
vapors will be discussed along with the significance of these results for
methods of fire debris sample analysis
Fire Debris, Ignitable Liquids, Trace Evidence
B101 Solid-Phase Microextraction Based
Approach for Enantiomeric Analysis
of Amphetamines
Sheng-Meng Wang, PhD*, Central Police University, 56 Shu-Ren
Road, Kuei-Shan, Graduate Program in Forensic Science, University
of Alabama at Birmingham, Birmingham, AL, Taoyuan, 333, Taiwan;
Ray H. Liu, PhD, Department of Justice Sciences, University of
Alabama at Birmingham, Graduate Program in Forensic Science,
Birmingham, AL 35294-2060
The goal of this presentation is to advance the development in
solid-phase microextraction (SPME), specifically, to explore an one-step
absorption/derivatization approach for enantiomeric analysis of amphetamines.
This presentation will benefit the forensic scientist’s analytical
knowledge of these SPME techniques.
As a solventless approach and with advances in fiber manufacturing technology, SPME has great potential in various sample pretreatment processes. Reported applications of this technology to the
analysis of amphetamines included: (a) the inclusion of the derivatizing
reagent in the sample matrix [1] or the chromatographic injection port
[2]; (b) a two-step approach in sequentially placing the fiber in the headspace of the sample- and derivatizing reagent-containing vials [3]; and
(c) an one-step procedure in placing the fiber in the headspace of the
derivatizing reagent-containing vial, which was in turn placed in the
headspace of the sample vial [4]. This current study represents a further
progress in the application of the SPME technology. Specially, an onestep process is used to complete the absorption/derivatization process for
the analysis of the enantiomeric compositions of amphetamines.
(S)-(-)-N-(Trifluoroacetyl)-prolyl chloride (l-TPC) was adapted as
the chiral derivatizing reagent and added directly into the sample matrix.
Temperature, absorption/desorption duration, and the amount of derivatizing reagent were studied to determine their effects on the yields of
analytes on the fiber. The derivatization products resulting from this
study show excellent desorption characteristics of the polydimethylsiloxane-coated fiber (100 µm) used in this study. For example, an onetime 5-min desorption leaves no detectable carry over. Optimal operational parameters (absorption: 70 oC for 10 minutes; injection: 250 oC
for 5 minutes) cause minimal negative impact on the fiber, allowing
repeated use (> 30 times) of the fiber.
This method was evaluated for its effectiveness (a) in quantitative
determination of the enantiomeric pairs of amphetamine and methamphetamine — in terms of repeatability, linearity, and limits of detection
and quantitation; and (b) by comparing its analytical findings with those
derived from a conventional liquid-liquid extraction approache
(Table 1).
* Presenting Author
Table 1. Comparison of methamphetamine/amphetamine enantiomeric
compositions resulting from two sample preparation protocols (SPME
and liquid-liquid extraction)
SPME (concentration in ng/mL)
Sample d-Methamp l-Methamp
d-Amp
l-Amp.
1
2
3
4
5
6
7
8
9
10
11
12
2,477
1,010
1,329
2,581
2,675
3,324
1,466
580
1,971
2,376
2,220
1,308
72
42
36
94
93
92
78
403
50
84
78
90
12,169
1,821
7,436
13,845
3,766
10,745
3,090
2,482
5,398
6,667
6,761
8,600
1,280
219
593
1,960
340
1,254
704
5,863
486
608
947
972
Liquid-liquid (concentration in ng/mL)
Sample d-Methamp l-Methamp
d-Amp
1
10,638
1,564
2,702
2
1,427
146
976
3
7,768
429
1,318
4
11,753
1,233
2,545
5
3,310
252
2,763
6
9,931
1,315
2,973
7
2,564
591
1,381
8
2,010
6,317
482
9
4,680
435
1,928
10
5,649
543
2,230
11
5,632
449
2,100
12
6,276
950
1,276
l-Amp.
63
32
37
67
72
79
79
538
43
66
73
58
References
1. H.G. Ugland, M. Krogh, and K.E. Rasmunssen. Aqueous alkylchloroformate derivatization and solid-phase microextraction: Determination
of amphetamines in urine by capillary gas chromatography. J.
Chromatogr. B 701: 29-38 (1997).
2. N. Nagasawa, M. Yashiki, Y. Iwasaki, K. Hara, and T. Kojima. Rapid
analysis of amphetamines in blood using head space-solid phase
microextraction and selected ion monitoring. Forensic Sci. Int. 78: 95102 (1996).
3. C. Jurado, M.P. Gimenez, T. Soriano, M. Menendez, and M. Repetto.
Rapid analysis of amphetamine, methamphetamine, MDA, and MDMA
in urine using solid-phase microextraction, direct on-fiber derivatization,
and analysis by GC-MS. J. Anal. Toxicol. 24: 11-16 (2000).
4. M.-K. Huang, C. Liu, and S.-D. Huang. One step and highly sensitive
headspace solid-phase microextraction sample preparation approach for
the analysis of methamphetamine and amphetamine in human urine.
Analyst 127: 1203-1206 (2002).
SPME, Enantiomeric Analysis , Amphetamines
* Presenting Author
B102 Scientific Validation of the Reliability of
Detector Dog Teams: Progress Towards
the Development of a National Standard
for Best Practices
Douglas P. Heller, PhD, MBA* and José R. Almirall, PhD,
International Forensic Research Institute, Florida International
University, University Park, Miami, FL 33199; Stefan Rose, MD,
University Medical & Forensic Consultants Inc, 10130 Northlake
Boulevard, Suite 214, #300, West Palm Beach, FL 33412; Kenneth G.
Furton, PhD, International Forensic Research Institute, Florida
International University, University Park, Miami, FL 33199
This presentation highlights recent progress towards developing
scientifically sound best practice procedures validating the reliability of
detector dog teams. Current scientific research and draft best practices
developed at biannual national detector dog conferences are presented
This presentation will update interested parties on recent progress
towards developing scientifically sound, best practice procedures validating the reliability of detector dog teams.
Even with technological advances in instruments, detector dogs still
represent one of the most reliable and widely used real time detectors of
contraband and forensic evidence. Unfortunately, to date, there have
been limited peer-reviewed published scientific studies demonstrating
exactly how these biological detectors work so efficiently and limited
scientific assistance to improve performance and confirm the stated reliability of these detection teams. Recent criminal investigations focusing
on bomb dogs have highlighted the need to have a standardized method
for confirming the performance of detector dog teams.
This poster discusses the critical and independent evaluation of the
selectivity and sensitivity of law enforcement detector dog teams and the
refinement of optimal methods to validate the performance of these
teams. The data indicate that canine detection teams that undergo sufficient initial training and maintenance provide scientifically reliable
detection of suspect samples with high precision and accuracy. A model
scientifically sound certification protocol for detector dogs including
drug and explosive canines has been evaluated. Important training and
maintenance factors include the use of double blind tests, positive controls minimizing contamination issues and negative controls including
representative distractors.
Also discussed are the results of the refinement of draft best practices for detector dog teams at the 3rd National Detector Dog Conference
held recently in North Miami Beach, Florida. Highlights of the group
discussions included the need for standardization of nomenclature, specific recommendations based on the work function of the detection team
(narcotics, explosives, etc.), general requirements for detector dog
trainers, protocols for certification of detector dog teams and best practices for maintenance and deployment record keeping. As one example,
certification should only be valid for the specific handler/dog team,
should last for one year, and requires that the handler conduct regular
maintenance training, among other requirements.
Towards the pursuit of a National Standard for detector dog team
best practices, the authors are in the process of initiating a planning
panel to explore establishing a Technical Working Group in conjunction
with the Department of Homeland Security’s Transportation Safety
Administration and other national authorities in the field. The best practices draft document should serve as a resource towards the development
of a National Standard. The beneficiaries of such a reference standard
may include practitioners and policy makers at federal agencies, law
enforcement and the legal and judicial community, and those in the
private sector who work with detection canines. In addition, if there is
to be a National Standard, challenges will be in bridging gaps with other
agencies and detection canine associations with respect to their existing
Standards and protocols, the harmonization of nomenclature being but
one of potentially several issues.
73
Scientific determination of the reliability of detection teams and the
use of scientifically validated teams also facilitates scientific research
aimed at identifying the active odor signature chemicals used by canines
and the development of improved instrumental methods capable of both
particle and vapor detection. Examples from the authors’ labs applied to
drug, accelerants and explosives detection will be highlighted.
Best Practices, Detector Dogs, Narcotics and Explosives
B103 Continued Scientific Investigation
of Explosives Detection by Canis Lupis
var. familiaris
Ross J. Harper, MSc* and Farzan Khodadadi, International Forensic
Research Institute, Florida International University, Department of
Chemistry and Biochemistry, Miami, FL 33199; Stefan Rose, MD,
University Medical & Forensic Consultants Inc, 10130 Northlake
Boulevard, Suite 214, #300, West Palm Beach, FL 33412; José R.
Almirall, PhD, and Kenneth G. Furton, PhD, International Forensic
Research Institute, Florida International University, Department of
Chemistry and Biochemistry, Miami, FL 33199
After attending this presentation, attendees will have a better understanding of canine detection best practices.
This presentation will provide a better understanding of canine
detection best practices. Acceptance of the differences between canine
training with real explosives or simulated training aids and the impact
that this can have on the quality of the canine performance.
This presentation will communicate the continuation of previous
work by the authors, focused upon identification of the active odour signatures of explosives, detailing the new results and broadening the scope
of samples studied to included explosives not previously presented. With
a view to better understand and give scientific validation to biological
detection of explosives by detector dogs, this work continues with more
odours identified and currently under investigation.
Another area of work appropriate to this study is the diffusion of
explosive odors though a variety of polymers. It is proposed that
through appropriate choices of plastic material, the target odor may be
released at a controlled rate to produce a highly effective training aid.
Gravitometric studies into the loss of explosive material from polymer
packaging are also presented in this paper.
The analysis and characterization of the headspace ‘fingerprint’ of
a variety of explosives, followed by canine trials of the individual components to isolate and understand the target compounds that the dogs
alert to. Studies to compare commonly used training aids with the actual
target explosive have also been undertaken to determine suitability and
effectiveness. The reliability of commercially available canine training
aids is investigated
The vapor headspaces of a range of explosives have been collected
using Solid Phase Micro Extraction (SPME) and analyzed by chromatography-mass spectrometry. Using SPME-GC-MS, samples of a
variety of explosives obtained from local law enforcement agencies for
odor determination have been characterized, and the prevalent signature
odor chemicals identified. Studies have also been performed using
SPME-HPLC-MS to observe the non-volatiles, and thermally unstable
compounds within the samples.
Following the characterization of the potential odor signatures of
the explosives, canine detection is then applied to a combination of
double-blind trials of the individual components from the odor signature.
The chemicals identified by canine trial are then packaged in a variety of
polymer bags, as prototype designs for safe, inactive training aids
SPME has been demonstrated to have a unique capability for the
extraction of volatiles from the headspace of explosives. Results to date
have shown comparable, chemical differences between readily available
74
training aids and the actual explosive matrices that they seek to replicate.
Odor signatures of TNT, RDX based plastic explosives, and assorted
propellants are presented.
Current work, including recent dog trials, has confirmed the
hypotheses that 2,4-DNT and 2-Ethyl-1-hexanol are the active odor signatures of TNT and C-4/Plastic Explosives respectively. Dog trials have
also discounted the taggant DNDMB, and diphenylamine as previously
proposed active odor signatures for C-4/Plastic Explosives and
Smokeless Powder respectively. Investigation into the effectiveness of
commercially available training aids has highlighted several major concerns regarding the suitability of the aids, and their simulated odors,
regarding canine training.
Canine Detection, Explosives, SPME-GC-MS
B104 Comparison of Field Portable Instruments
and Accelerant Detection Canines for the
Detection and Collection of Ignitable
Liquid Residues at Fire Scenes
Shirley S. Chin, BS* and Kenneth G. Furton, PhD, International
Forensic Research Institute, Florida International University,
University Park, Miami, FL 33199
This presentation will demonstrate a complementary method used
in the search for ignitable liquid residues (ILR) at fire scenes when
Accelerant Detection Canines (ADCs) are not available, or when the
hazards of the scene prevent the use of such dogs, while maintaining the
sensitivity and selectivity of ADCs. In addition, an investigation into the
use of an on-site portable ILR sampler for the separation and extraction
of volatile accelerants will be discussed. It is proposed that this method
would hopefully simplify the collection and transportation of samples
from fire debris for subsequent chromatographic analysis in the laboratory.
This presentation will demonstrate the practicality of using these
portable detectors for fire/arson investigators to detect the use of accelerants at a fire scene, and to collect adequate samples for submission to
a forensic laboratory for analysis.
Arson, the intentional burning of property, is a serious crime in
many countries including the United States. The use of flammable
liquids is a frequent choice for arsonists to help accelerate the spread of
a fire. One of the responsibilities of the fire investigator is to look for
signs of accelerant use that could have started these incendiary fires.
Chemical analysis of accelerant residues in the fire debris would assist
the investigator in determining if the fire was due to intentional or
natural causes.
It is well known that dogs have superior olfactory capabilities, thus
Accelerant Detection Canines (ADCs) have been employed at some fire
scenes to pinpoint locations where flammable liquids had been used. It
has been reported that ADCs can distinguish between some accelerants
and interfering pyrolysis products of common materials found at the fire
scene. Not only are they able to discriminate between the accelerants and
pyrolysis products, but it has also been reported that ADCs can detect
down to levels below the sensitivity limits of analytical instruments used
in forensic laboratories. Although accelerant detection canines have been
involved in court cases in the United States, there have also been controversies over admissibility of canine testimony. Unless confirmed by
laboratory analysis, canine alerts would raise a reliability issue in court.
A complement to using these biological detectors is to use electronic sniffers. The purpose of this study is to determine the capability of
one such on-site electronic detector compared to ADCs. The TLV
Sniffer® (Scott Instruments) is used to detect the presence of combustible gases. This instrument was put through similar proficiency tests
that accelerant detection canines perform, as outlined by the Canine
Accelerant Detection Association (CADA).
* Presenting Author
Since the TLV Sniffer® is designed to detect and measure concentrations of combustible gases, it will also indicate the presence of hydrocarbons from pyrolysis products. For confirmatory purposes, another
field instrument was considered. The Portable Arson Sampler (PAS) separates and collects volatile accelerants from the fire debris. Ideally, one
would use the TLV Sniffer® to first locate the source(s) of accelerants
among the fire debris. Once located, the PAS would be used to sample
the headspace of that fire debris and extract the volatile gases onto
adsorbent tubes. The contents of the tubes would then be chemically or
thermally desorbed for subsequent gas chromatography-mass spectrometry (GC/MS) analysis at the laboratory.
To study the selectivity of the TLV Sniffer®, various substances
that could be found at a fire scene, including pinewood, Styrofoam,
newspaper, and nylon carpet, were set on fire. Comparisons between
fires that were extinguished by oxygen deprivation and by dousing with
water were carried out. Headspace analyses of these burned matrices
were conducted, both with and without being spiked with 5uL 50%
weathered gasoline.
Results show that the weathered gasoline can still be detected under
both extinguishment conditions, even after air venting for 30 minutes.
Burnt wood, nylon carpet, and Styrofoam produce pyrolysis products,
but these would not be confused with gasoline once analyzed by
GC/MS. However, burned black and white newsprint paper generates
common compounds also present in gasoline. But to a skilled analyst,
their chromatograms would not be mistaken due to the ratios and patterns of the peaks generated by gasoline. A problem may arise, however,
when the sample has been air vented for 30 minutes or more, as most of
the alkylbenzenes of gasoline will have evaporated. Thus there is a need
for adequate comparison samples to act as a negative control. Overall,
these results indicate that electronic sniffers and adsorption samplers can
be useful complements to ADCs, as they offer an objective qualitative
analysis which can be used in a court of law. In addition, there are advantages of the on-site sampling capabilities offered by the PAS including
reduction to loss of sample, less bulky packaging sent to the laboratory,
and the ability to sample from areas that cannot be conveniently transported to the laboratory.
Arson Investigation, Electronic Detectors, Accelerant Detection
Canines
B105 National Forensic Laboratory Information
System: 2003 Data Analysis
Michael R. Baylor, PhD*, Research Triangle Institute, 3040 Cornwallis
Road, PO Box 12194, Research Triangle Park, NC 27709-2194;
Linqun Wong, MS and Frank L. Sapienza, MS, Drug Enforcement
Administration, Office of Diversion Control, 600 Army Navy Drive,
Arlington, VA 22202; Kevin J. Strom, PhD and Valley Rachal, MS,
Research Triangle Institute, 3040 Cornwallis Road, PO Box 12194,
Research Triangle Park, NC 27709-2194
After attending this presentation, attendees will have an understanding of the National Forensic Laboratory Information System
(NFLIS) that systematically collects results from drug analyses conducted by state and local forensic laboratories. The data and results presented will provide a detailed approximation drug evidence seized by
law enforcement agencies and sent to forensic laboratories for analysis
from January 1, 2003 to June 30, 2003.
This presentation will provide timely data on the variation in the
distribution of controlled substances across geographic areas, identify
emerging drugs of abuse identified in forensic laboratories, provide
information on the diversion of licit drugs into illicit channels, and
improve the ability to estimate drug availability based on laboratory
analysis and identification.
This poster will describe the NFLIS program which first began
implementation by the DEA and Research Triangle Institute (RTI) in
* Presenting Author
September 1997. To date, approximately 36 state forensic laboratory
systems and 56 local forensic laboratories that perform drug analyses,
comprising a total of 192 individual forensic crime laboratories
throughout the United States, have joined NFLIS. With the goal of
including all of the forensic laboratories in the United States, the sustained recruitment of non-participating state and local forensic laboratories remains a priority.
NFLIS provides results of drugs analyzed and reported by participating labs through semi-annual reports, annual reports, and the
Interactive Data Site (IDS). The IDS combines timely and detailed data
analyses with a flexible, user-friendly system. It allows participating
laboratories to run parameterized queries against the NFLIS database in
a near real-time capacity. Labs can initiate queries for their own data at
the individual case-level or they can calculate aggregate regional and
national results. IDS users can specify the time period, region, type of
laboratory, and drug type in order to customize these queries. Aggregate
results of drugs identified and reported by participating NFLIS laboratories representing the period January 2003 through June 2003 will be
presented. Highlighted findings will include the estimated prevalence of
selected “drugs of interest” and analyzed drug items by category. The
distribution of drug items by percent and number of total analyzed items
in the state and local forensic laboratories will be depicted. The number
and percentage of analyzed drug items for the twenty-five most frequently reported drugs, as well as the major drug categories such as narcotic analgesics, benzodiazepines, “club drugs,” stimulants, and anabolic steroids will be presented in tables and graphs. Special study data
on (1) drug combinations, (2) drug purity, and (3) drugs identified in
strategic geographic locations as well as major metropolitan areas will
be summarized.
The NFLIS data base is serving the drug enforcement community:
supporting drug control/drug scheduling, highlighting variations in distribution of controlled substances across geographic areas and over time,
improving estimates of drug availability, providing timely information
about the diversion of licit drugs into illicit channels, identifying
emerging drugs of abuse, increasing our understanding of the nation’s
drug problem, and linking the drug enforcement and forensic laboratory
community across the nation.
Drug Analysis, National Forensic Laboratory Information System,
Drug Database
B106 Elemental Analysis of Materials by Laser
Ablation Inductively Coupled Plasma
(LA- ICP-MS) for Forensic Applications;
Instrumental Considerations
José R. Almirall, PhD* and Tatiana Trejos, BSc, Florida International
University, Department of Chemistry, University Park, Miami, FL
33199
This presentation will cover the instrumental considerations associated with the analysis of trace amounts of metals in materials such as
differences between sample introduction systems, mass spectrometers
and available options.
This presentation will facilitate a better understanding of the use of
the different sample introductions systems into an ICP-MS. The presentation will also cover the selection of spectrometer types for the purpose
of trace elemental analysis, including the use of TOF, quadrupole and
magnetic sector instruments. An analysis of the dynamic reaction cell
technology as applied to forensic analysis, is also presented.
The elemental analysis of materials has become an important yet
underutilized type of evidence at many crime scenes. Although the
utility of trace elemental analyses and comparisons for glass or paint
fragments has been shown to offer a high degree of discrimination
between different sources of these materials, the lack of method devel75
opment, validation of methods and publication of results in the open literature have limited the adoption of this technology by the typical
forensic laboratory. Additional barriers include the expense of the
equipment and the sophistication required for its operation.
This presentation will focus on presenting the commercially
available options for the elemental analysis of materials such as glass
using ICP-MS, the most sensitive and practical of the elemental analysis
methods available to the forensic scientist. Forensic examiners have the
option to analyze glass by dissolution (digestion) or, by using a solid
sample introduction system (laser ablation). The advantages and disadvantages of each of these choices are presented. Forensic examiners then
have the option of selecting the type of mass spectrometer as the
detector. Time of Flight (TOF), quadrupole and magnetic sector instruments are available. Speed of analysis, resolution and cost considerations will determine the instrument of choice and these factors are discussed. Within the quadrupole class of mass spectrometers, new collision cell accessories are now available in order to reduce the interferences that often plague unit resolution devices.
While solution analysis does not require the additional purchase of
a laser ablation system, significant disadvantages related to the digestion
of glass samples leads one to conclude that the added expense of the
solid sampling introduction system is worthwhile. A direct comparison
of solution data with LA-ICP-MS data for a variety of glass sample types
is presented. A direct comparison of the data generated using the different mass spectrometer detectors is also presented. Finally, a study to
determine the utility of a dynamic reaction cell using a Perkin Elmer
DRCII mass spectrometer as compared to the use of an Agilent 4500
plus mass spectrometer is also presented. The most significant advantage
to the addition of dynamic reaction cell technology is the potential for
the elimination of polyatomic ion interferences for Se, As, Fe, K, Cr, Mn
and Cu upon detection.
A direct comparison between two laser systems (CETAC LSX 200
plus and the CETAC LSX 500) when used for materials analysis is also
presented. The LSX 500 offers better limits of detection and precision
(<5% RSD) due to the increased power of the laser and the flat beam
profile, both of which lead to better laser/material interaction. Scanning
Electron Microscopy (SEM) imaging was also conducted in order to
evaluate physical characteristics of the craters left by the laser in the
glass fragments. A JSM-5900-LV JEOL SEM operated at high vacuum,
using secondary electron imaging at 20 KV was used to image craters of
a spot size of ~ 40µm. The glass samples were coated with gold to
prevent charging.
succumbed to a seven year battle with cancer two weeks prior. Several
questions required an answer: Who was the child? When and how did
the child die? Why were the remains in the locker? Had a crime been
committed?
Due to the mummification of the remains, the sex of the child and
the cause of death could not be determined at autopsy. The question of
identity was answered by DNA profiling of the bone marrow from the
left and right femurs of the infant. DNA typing was performed using the
AmpFlSTR® Profiler Plus and COfiler Amplification Kits, and a 13
STR loci female DNA profile was obtained. The DNA profile of the
condominium owner proved elusive, and attempts were made to obtain
it from personal effects, as well as two bladder biopsy samples. The full
profile was eventually determined by analysis of a bloodstain in a pair of
underwear that had been worn at the hospital during the last few days of
her life. It was determined that she could not be excluded (LR =
520,000) as the biological mother of the infant. The adopted daughter, on
the other hand, was excluded.
The question of when the child died remained unanswered. A U.S.based expert estimated that mummification had taken approximately 69
days, based on an analysis of the infant’s clothing and an assessment of
various factors in the decomposition of human remains; however, this
information was of no assistance in establishing the time of death. The
condominium owner’s ongoing medical treatment for the past seven
years meant that it was not likely that the infant could have been born
during this period of time. Another clue to the year of the birth was provided by an assessment of the age of the diaper. Photographs of the
diaper were sent to a manufacturer (Kimberly-Clarke) and it was determined that the particular style had been discontinued in 1984.
With the time frame narrowed down to the mid 1980s, investigators
questioned the relatives, friends and work colleagues of the owner as to
who the father could have been. None of these individuals had any idea
that she had been pregnant at any time; however, the identity of a
potential father was determined. The ex-boyfriend, who had moved to
England since their involvement, had no knowledge of the pregnancy
and did not think that the infant could be his. DNA profiling was carried
out and it was determined that the ex-boyfriend could not be excluded
(LR = 61,000) as the biological father of the mummified child, and the
year of the birth was thus established as the year in which the relationship ended, 1985.
No cause of death could be established, due to the condition of the
body, and therefore no charges were laid.
Mummification, Familial DNA, Major Case Management
Elemental Analysis, LA-ICP-MS, Materials Analysis
B107 The Mystery of the Mummified Baby
Alison Morris, MSFS* and Roger Frappier, MSc, Centre of Forensic
Sciences, 25 Grosvenor Street, Toronto, ON M7A 2G8, Canada; Jim
Cairns, MD, Office of the Chief Coroner, 26 Grenville Street, Toronto,
ON M7A 2G8, Canada; Mike Stoker, Toronto Police Service, 52
Division, 255 Dundas Street West, Toronto, ON M5T 2W5, Canada
This presentation will highlight a multidisciplinary investigation
performed over a two year period in a case from Toronto, Ontario, that
involved the Office of the Chief Coroner, the Toronto Police Service,
and the Centre of Forensic Sciences, working together to manage a
major case from both the scientific and investigative perspectives.
This general interest presentation, involving different scientific disciplines, including forensic pathology and biology, will demonstrate the
value of working in partnership to solve a challenging case.
On July 16, 2001, the mummified remains of an infant were discovered in the storage locker of a Toronto waterfront condominium by
the 150-year-old adopted daughter of its female owner. The infant was
found inside a garbage bag, clothed in a diaper and shirt, and wrapped in
blankets. The condominium owner, a successful business woman, had
76
B108 Cat or Dog Hair?—The Root
of the Problem
Agnieszka Janusz, BSc, Forensic Science Centre, 21, Divett Place,
Adelaide, 5000, Australia; Silvana Tridico, BSc*, Centre of Forensic
Sciences, 25 Grosvenor Street, Toronto, ON M7A 2G8, Canada
After attending this presentation, attendees will learn that the differentiation between cat and dog hairs based solely on the appearance of
a spade-shaped root, cited in standard texts as being a characteristic of
dog hairs, may lead to an erroneous identification.
The presentation will demonstrate that the identification of putative
cat or dog hairs based solely on the appearance of a spade shaped root
may result in an erroneous identification.
This poster will present results of a study conducted at the Forensic
Science Centre, which shows that cat hairs may possess spade shaped
roots comparable to the ones typically found on dog hairs. The differentiation between the spade shaped roots of these two animal species may
be made on the basis of their lengths.
The present study was initiated as a result of findings during the
examination of a large number of animal hairs related to a double
homicide case in Adelaide, South Australia. During the microscopic
* Presenting Author
examination of these hairs the author identified the majority of the hairs
as being dog in origin. However, during the course of the examination a
few animal hairs although bearing spade shaped roots exhibited other
features characteristic of cat hairs. The spade shaped roots on the
putative cat hairs were significantly shorter than the spade shaped roots
on the hairs identified as originating from a dog. This finding prompted
the author to conduct a preliminary study to determine whether cat hairs
possessed spade shaped roots and if so, to measure the lengths.
Guard hairs from 21 dogs and 20 cats were collected by grooming
an animal, either by hand or with a brush, along the entire length of its
back. Some smaller breeds of dogs were deliberately chosen in addition
to larger ones in order to determine if the spade root characteristic
lengths varied with the size of breed.
Guard hairs were chosen because these hairs exhibit characteristics
that are the most useful in identifying the animal of possible origin. No
attempt was made to differentiate between hairs from different parts of
the body, as the forensic scientist is not usually concerned from which
part of an animal’s body a particular hair originated.
Ten hairs were selected at random from each sample, resulting in a
total of 411 hairs (199 hairs from cats and 212 from dogs). The unequal
number of hairs for each animal was due to some of the hairs not bearing
roots. Each of the ten hairs was individually mounted in XAM mounting
medium, on labeled microscope slides. The roots were examined on a
compound, transmitted light microscope capable of up to 400x magnification. The shapes of each root and their lengths were noted. The lengths
were determined in microns using an eyepiece graticule calibrated
according to the manufacturer’s instructions.
The examination of the cat hairs revealed that approximately 30%
of the hairs examined exhibited a spade root comparable to the ones seen
on the dog hairs. The spade-shaped cat hair roots were not limited to any
particular breed of cat. The most significant difference between the
spade roots of the two genera was length. The average length of the cat
spade roots was approximately 200 microns, whilst those of the dog
spade roots were approximately 500 microns. An overlap region of 260320 microns occurred between the respective roots of the animals.
The results of this study highlight that whilst the standard works
serve as excellent guidelines to the identification of animal hairs it is
crucial that the hair examiner be aware that they are not definitive or
exhaustive works. The results of the study indicate that if the determination on whether a putative hair is of cat or dog origin is based solely
on the appearance of a spade root an erroneous identification may be
made.
Spade-Shaped Root, Forensic Animal Hair Analysis, Animal Hair
B109 Optimization of Collection and Storage
Methods for Scent Evidence and the
Identification of the Volatile Components
Comprising an Individual Human Odor
Signature
Allison M. Curran, BS* and Kenneth G. Furton, PhD, Florida
International University, Chemistry Department, CP-345 University
Park, Miami, FL 33199
After attending this presentation, attendees will understand the best
procedure for the collection, as well as the preservation, of human scent
evidence samples and different passive headspace sampling techniques
used to evaluate the uniqueness of human odor.
humanity by demonstrating the optimization of scent evidence collection
and preservation procedures as well as an evaluation of the uniqueness
of human odor may lead to scent evidence being utilized in the U.S. as
an investigative tool in criminal cases.
* Presenting Author
The use of canines, Canis lupus var. familiaris, within the fields of
law enforcement and forensic science is widely accepted, however, to
date, there are few peer reviewed studies confirming their accuracy and
reliability. The use of detector dogs for “human scent lineups” has been
utilized in European countries (such as the Netherlands) but has not
gained widespread acceptance in the United States due, in large part, to
the lack of definitive studies demonstrating the reliability of this
approach. This lack of information has resulted in successful legal challenges to the use of these biological detectors in a court of law.
Fingerprints recovered from evidence and at crime scenes can be
used to identify suspects. However, if a print recovered from a crime is
smudged or incomplete it may be hard to distinguish between individuals in question. The identification of the chemical and volatile components of the print may aid in the discrimination between individuals in
question. The identification of the chemical components within fingerprint residue may lead to a ‘chemical fingerprint’ of the individual as
may also hold true for the volatile components of fingerprint residue.
These volatile components are what comprise the signature odors that
law enforcement certified canines alert to when searching for humans
and distinguishing between individuals.
Chemical residues of human fingerprints have been studied in some
detail but, at present, there have been few reports of the volatile constituents of human odor from various parts of the body. The following
diagram shows some of the chemical components found in fingerprint
residue:
This paper describes the use of headspace extraction combined with
gas chromatography and mass spectrometry to identify the signature
odors that law enforcement certified detector dogs alert to when
searching for humans and distinguishing between individuals.
Comparisons of two different passive headspace extraction techniques:
solid phase micro-extraction (SPME) and activated charcoal strips
(ACS) will be presented. Studies include the analysis and identification
of the headspace ‘fingerprint’ of a variety of samples, followed by completion of double-blind dog trials of the individual components in an
attempt to isolate and identify the target compounds to which dogs alert.
SPME–GC/MS and SPME-LC/MS have been demonstrated to have
unique capabilities for the extraction of volatile compounds from the
headspace of forensic specimens and show great potential to aid in the
investigation and understanding of the complicated process of canine
odor differentiation. Major variables evaluated for the headspace SPME
include fiber chemistry, a variety of sampling times, and the resultant
effect on ratios of isolated volatile components.
Various methods for sampling human scent will be presented,
including the use of the Scent Transfer Unit – STU-100 (Lynn Peavey
Company, Lenexa Kansas). Different mediums are being evaluated for
use in the collection of human scent, such as: sorbents (silica, octadecyl,
Florisil, etc.), cotton pads (soft, stiff), cotton balls, polyester, and wool.
77
Sampling methods and mediums are being investigated in an attempt to
optimize the recovery and storage of human scent from forensic specimens. Persistence/dissipation studies will also be presented evaluating
variables such as static conditions, dynamic airflow, and temperature
effects. Various containers are being evaluated for use in storage of
human scent samples, including: glass jars, paint cans (lined, unlined),
bottles, Ziplock bags, heat sealed bags (i.e., Kapak), as well as different
plastics. Headspace and bulk human scent residues collected in different
ways will be compared. Determining the effect of odor traces collected
from different areas of the body (feet, hands, underarms, etc.) will also
be presented.
Canine, Human Scent, Odor Signature
B110 LC/MS Analysis of Flunitrazepam
(Rohypnol®) Solid Dosage Tablets
Andrew Benson, MS, Florida International University, Department of
Chemistry, University Park CP-178, Miami, FL 33174-3509; Stefan
Rose, MD, Florida International University, University Park CP-178,
Miami, FL 33199; Yali Hsu, MS, Florida International University,
Tamiami Park CP-178, Miami, FL 33199; Kenneth G. Furton, PhD,
Florida International University, University Park, Department of
Chemistry, DM-445, Miami, FL 33199; Alberto Sabucedo, PhD*,
Florida International University, Department of Chemistry, Miami,
FL 33199
qualitative and quantitative analysis of flunitrazepam by Liquid
Chromatography Mass Spectrometry (LC/MS). This poster will convey
the process of extracting and quantitating this drug by LC/MS. It
includes quantitation and confirmation by LC/MS.
This presentation will demonstrate to the forensic community that
LC/MS will provide an improved methodology over GC/MS methods
related to the analysis of this drug. This chromatograpy and sensitivity
is a significant improvement over current GC/MS techniques.
The quantitative analysis of seized flunitrazepam tablets can be performed using liquid chromatography-mass spectrometry (LC/MS).
LC/MS is ideal because of the physical properties of flunitrazepam and
the faster analysis times relative to gas chromatograph-mass spectrometry (GC/MS). Gas chromatography-mass spectrometry (GC/MS)
techniques are not ideal for molecules that are polar and have low
volatility such as flunitrazepam. Liquid-liquid extraction (LLE) was
used to isolate flunitrazepam from the inert water-soluble constituents of
the tablet. The technique is rapid, sensitive and confirmatory for flunitrazepam in solid dosage tablets.
Flunitrazepam is a class of potent hypnotic agents available on prescription in most European and Latin American countries. In the United
States flunitrazepam is a scheduled drug of abuse. Many date-rape cases
have been linked to the incapacitating effects of flunitrazepam. These
effects are exacerbated by the presence of alcohol typically found in
environments where flunitrazepam is illegally utilized. The availability
of these pills originates from the illegal smuggling from countries that
manufacture this drug for therapeutic purposes.
The tablets are dissolved in water to break up the tablet matrix. The
tablets form an aqueous suspension that is mixed with an equal volume
of chloroform. The sample is vortexed and sonicated for a few minutes.
The chloroform layer is allowed to separate by centrifugation for a few
minutes. The denser chloroform layer is aspirated with a pipette to avoid
contamination of the organic phase with the aqueous phase. The procedure should be repeated five times to be able to achieve quantitative
recovery of flunitrazepam. It should be noted that basic extractions
should be avoided because of the instability of flunitrazepam in alkaline
medium. The higher the pH the faster the degradation kinetics of flunitrazepam. We used distilled water instead of an alkaline aqueous solution
to perform the liquid-liquid extraction.
78
The assay used a deuterated internal standard, flunitrazepam-d7, to
correct for changes in running conditions. A C-18 reverse phase column
was used to 4.6 x 150mm, 1 mL/min, methanol:water (70:30) mobile
phase with isocratic elution. A plot of the ratio of the areas of
[Flunitrazepam]/[Flunitrazepam-d7] versus the respective concentration
gave us an excellent linear regression fit for the standard curve. The linearity range of the assay was 0 to 500 ng/mL. The standard linear
regression line had a coefficient of correlation, r = 0.999. The limit of
detection (LOD: S/N =3) was 12 ng/mL (injection volume 20uL) and the
limit of quantitation (LOQ: S/N = 10) was 50 ng/mL. The assay was run
in selected ion monitoring mode (SIM) choosing several ions that can be
used for qualifying and quantitation. The need to produce fragmentation
can be achieve by increasing the cone voltage of the Finnigan aQa
Navigator™ LC/MS system. This is important for forensic work where
a minimum of three ions, ion ratios within +/-20%, and retention times
are part of the confirmatory requirements for selected ion monitoring.
The ions monitored for flunitrazepam were m/z 314, 300, 272, 239
and for the deuterated internal standard, flunitrazepam-d7, m/z 321, 307,
279, 246. The highest sensitivity for this assay can be obtained with a
lower cone voltage, but the fragmentation pattern would not be
achieved. The analysis of solid dosage tablets is not usually dependent
on sensitivity of the assay because of the high concentrations of drug
found in the tablets.
The analysis performed on a confiscated pill presumably manufactured by Roche had a label concentration that read 1 mg. The tablet
appears to be legitimate based on its identifying marks. The quantitative
analysis by SIM mode LC/MS gave a recovery of approximately 1.08
mg flunitrazepam. The label number seems to indicate that this was a 1
mg flunitrazepam tablet. The assay showed quantitative recovery of flunitrazepam, recovery of tri-level controls were within 10% of target
value and our negative controls were negative. This indicates that our
deuterated standards did not contain detectable quantities of nondeuterated standard. In addition, no carryover effects were present at the
highest concentration of the linear range. The assay developed is a rapid,
sensitive and confirmatory for the presence of flunitrazepam in solid
dosage tablets. The extraction technique is simple, efficient and quantitative. It allows for a robust confirmatory technique for flunitrazepam by
LC/MS.
Flunitrazepam, Rohypnol®, Roofies
B111 Analysis of Explosives for Homeland
Security by Chemical Ionization GC/MS
Eric Phillips, BS*, Thermo Electron, 2215 Grand Avenue Parkway,
Austin, TX 78728
technique for analyzing low levels of explosives and residues.
Explosive compounds are by their very nature unstable. Analysis
of these compounds, precursors, and residues by GC/MS requires the
samples be forced into the gas phase without decomposition. This can
be accomplished by using a PTV injector. The sample is injected at a
temperature that is high enough to volatilize the solvent but not the compounds of interest. The injector temperature is then quickly ramped to
volatilize the compounds of interest. This injection technique also
allows larger sample volumes. This larger sample volume combined
with negative chemical ionization will allow extremely low detection
limits. The Trace DSQ eliminates neutral noise and there is virtually no
chemical noise when using negative chemical ionization. There are very
few types of compounds that will form a stable negative ion. Due to the
fast scanning rates of this instrument, fast GC techniques will be used.
EPA method 8095 includes compounds that are explosives,
explosive residues, and compounds that are used in the creation of
explosives. Following EPA method 8095 protocols, which is normally
performed on an ECD, the Trace DSQ with large volume injections and
* Presenting Author
negative chemical ionization is able to show a linear calibration curve
from 1 to 1000 pg/ul. This will show correlation coefficients of 0.99.
Scanning at 3000 amu/s will allow the GC run time to be less than 12
minutes and enough points across the peak for it to be fully characterized.
Homeland Security, GC/MS, Explosives
B112 Expansion of an AFLP DNA Marijuana
(Cannabis sativa) State, National, and
International Database
Eric J. Carita, BA*, Connecticut State Forensic Crime Laboratory, 278
Colony Street, Meriden, CT 06451; Heather Miller Coyle, PhD,
Department of Public Safety, Forensic Sciences Services, 278 Colony
Street, Meriden, CT 06451; Carll Ladd, PhD Timothy M. Palmbach,
MS, JD, and Henry C. Lee, PhD, Connecticut State Forensic Crime
Laboratory, Forensic Science Lab, 278 Colony Street, Meriden, CT
06451
The goal of this presentation is to demonstrate how AFLP can be
used to create databases of non-human origin and how they can be
applied to the forensic and law enforcement community.
This presentation will impact the forensic community by demonstrating how AFLP can be used to create databases of non-human origin
and how they can be applied to the forensic and law enforcement community.
Amplified Fragment Length Polymorphism (AFLP) analysis is a
polymerase chain reaction (VCR) based DNA typing method in which
amplification of restriction fragments are used to individualize single
source biological samples. The authors are currently in the latter stages
of completing state, national, and international AFLP marijuana
(Cannabis sativa) databases composed of law enforcement seizure
samples. The database has forensic significance in that it has the
potential to identify and link clonally propagated marijuana plants with
cultivators, distributors, and users as well as tracking certain “strains”
that illegally enter into and are circulated throughout the United States.
In order to determine the genetic variability of marijuana and its statistical correlation, a large marijuana population database is being created
from seizure samples provided by state and local authorities in
Connecticut, Vermont, Florida, Iowa, Kentucky, Wyoming, Tennessee,
West Virginia, Canada, and Taiwan. These seizure samples were used to
create state (100 samples), national (150 samples), and international
(200 sample) databases.
The database was created by using four selective primer sets
(EcoRT-ACT FAM/MseI-CAA, EcoRI-ACT FAM/MseI-CAT, EcoRlAAG JOE/MseI-CAT, and EcoRI-AAG JOE/Msel-CTA: Al, A4, F4, and
F5 respectively) from the Applied Biosystem’s AFLP™ Plant Mapping
Kit and separating PCR products by gel electrophoresis on an ABI 377
DNA Sequencer. 100 predetermined fragments are then scored in
Genotyper® (ABI) and converted to a binary code sequence that represents the samples genetic “profile.” This combination of “1s” and “0s”
are then imported into the database, which is used as a valuable search
tool for identifying samples that are consistent with clonality. However,
due to the fact that there is the possibility of two or more unrelated or
half-sibling samples being represented by the same binary code, samples
whose profiles match are then superimposed upon each other in
Genescan® (ABI) to determine whether any minor peaks outside the
defined categories are detected.
As with many forensic applications, statistics play an important role
when a database of this type is used in court. Two types of statistical
analyses are being conducted a) the counting method and b) the confidence interval. To determine the random match probability (RMP), the
conservative counting method of 1/N (N=the total number of unique
profiles in the database) is used. Since this statistical method may be
* Presenting Author
used, the greater the size of the database the more meaningful the statistical values will be. With this in mind, the authors are hoping to eventually create a 500 unique profile database. For an even rare conservative statistical approach two methods may be used. 1-a1-N (? =0.05 for
the 95%confidence interval) can be applied to profiles not previously
observed within the database, while for those profiles that have been
observed, P+/-1.96(vP(1-P/N) may be employed. Again, since a 95%
confidence interval is being used, the latter two statistical methods are
much more conservative than the more commonly used counting
method.
In demonstrating the usefulness of this technique and database, two
case study examples will be presented in detail.
AFLP, Marijuana, Population Database
B113 Case Report: Tangential Gunshot
Wound With MagSafe Ammunition
Amy V. Rapkiewicz, MD*, National Institute of Health Department of
Pathology, 10 Center Drive, Building 10-2N212, Bethesda, MD 20892;
Charles A. Catatnese, MD, Office of the Chief Medical Examiner, New
York City, SUNY/Downstate/Kings County Hospital Department of
Pathology, Brooklyn, NY; Robert Tamburri and Mark E. Basoa, New
York City Police Department, Firearms Analysis Section, 150-14
Jamaica Avenue, Jamaica, NY
a case report demonstrating the characteristic autopsy findings that can
be utilized to identify wounding patterns of unconventional prefragmented ammunition such as MagSafe ammunition.
This poster will present the wound characteristics that can be used
to identify MagSafe ammunition. MagSafe ammunition is a type of
unconventional prefragmented handgun ammunition that has great
wounding potential and therefore should be differentiated from other
types of prefragmented ammunition.
This case reports describes the fatal gunshot wound sustained by a
35-year-old woman. The unique features reported herein are the nature
of the wound and the ammunition used to produce it. The MagSafe bullet
produced a tangential type wound with an underlying keyhole defect of
the skull. The wound has an entrance defect that is contiguous with the
exit defect. The projectile disrupted the skin, subcutaneous tissue, bone
and brain matter leaving an elongated central defect in its path. The skin
defect is edged by lacerations, which are formed by skin tags. A skin tag
is defined as an elongated fragment of tissue connected to the margin of
a graze wound and projecting into its central portion. This injury is a
classic example of skin tag formation as a result of damage by a tangential projectile. The trajectory can be determined because the skin has
not been altered by decomposition, fire or surgical intervention. The lacerated skin has a semicircular abraded edge at the anterior aspect typical
of an entrance defect. The posterior aspect of the wound or exit end is
split. It should be noted that the lacerated border of the skin tag is the
edge toward the weapon and the abraded border adjacent to the skin tag
is away from the weapon. The lacerated edge will typically have
irregular margins and tissue bridging.
MagSafe Ammo is a type of prefragmented handgun ammunition.
Its construction consists of a copper jacket filled with birdshot that is
potted in a hard epoxy resin. MagSafe utilizes a poured epoxy cap to seal
the birdshot in the copper jacket. There are two distinctive features of
MagSafe ammunition: differential thickness of the epoxy cap and the
ability to choose the shot size. The poured epoxy cap can be made thin
for early fragmentation or thick for greater penetration. They are light
high-speed projectiles that deliver a huge release of kinetic energy and
fragment upon impact with a target. While this rapid transfer of energy
decreases penetration, it imparts remarkably devastating wounding
capabilities with less over-penetration and ricochet. The pieces of plastic
79
or epoxy can be used to identify this specific type of ammunition.
Gunshot wounds produced by this ammunition require careful examination in order to differentiate them from a shotgun or shot shell ammunition. There are many clues that can be used to accomplish this task.
In summary, this presentation is of a MagSafe gunshot wound to the
head with unique identifying features. Prefragmented bullets such as this
one are designed to release tremendous kinetic energy with decreased
penetration and ricochet. Differentiating types of prefragmented ammunition used to inflict gunshot wounds may be achieved by analysis of the
fragmented bullet components.
MagSafe Ammunition, Prefragmented Ammunition,
Tangential Wounds
B114 Mineralogical Changes in Soil —
A Great Tool to Forensic Explosive
Investigator
Bablinder K. Chopra, PhD*, Central Forensic Science Laboratory,
DFS, MHA, GOI, Hyderabad, Andra Pradesh 15, India; A.K.
Prasad, PhD, Punjab University, Sector 14, Chandigarh, UT 160014,
India; J.K. Sinha, PhD, House no. 1170, Sector 43 B, Chandigarh, UT
160043, India
The study has been done keeping in view the difficulties faced by
forensic experts in explosion cases. The presentation will impact the
forensic experts as the resume of conclusion of mineralological and geochemical changes occurring at the time of explosion due to tremendous
heat may be used in fingerprinting the explosive identification and time
lapse even after several years.
When a high explosive is detonated to cause explosion, it produces
a large quantity of gas and heat in a very short time resulting in development of a very high pressure. There is sustained shock wave and sufficiently effective blast pressure. During the explosion, the temperature
developed may be as high as 5000 ºC. There is absorption and adsorption
of volatile and nonvolatile constituents of explosive substances and their
combustion products in different layers of soil. Thus, the soil at the site
of explosion is likely to under go significant physical, chemical and mineralogical changes. Depending upon the type and quantity of explosives
there would be variation in the resultant changes. The larger the
explosive charge; the deeper will be the crater. The explosion will
accordingly affect even the deeper layers (B Horizon) of the soil
resulting in scattering of the affected soil to a larger area keeping the
crater as center. Therefore, the debris collected from the site of explosion
is usually available in large quantities. The first priority of examination
of soil samples so collected from the site of explosion is to detect and
identify the explosive used. A wide range of methods has been used by
workers in the field for analysis of post explosion residues. It may not
be possible to identify explosive materials if proper soil sample from the
debris is not available due to scanty amount of explosive substances
admixed with the soil. The explosive may not be identified from the
available small quantity of soil carried by the suspect from the site of
explosion in apparel, shoes and other belongings. It is still more difficult
after considerable time lapse. A foolproof methodology is therefore warranted for the examination of soil samples to arrive at some significant
forensic conclusions.
In view of the aforesaid discussion, a detailed study of post
explosion soil samples [exploded with RDX] has been undertaken and
mineralogical changes were studied by XRD and IR. A perusal of the
XRD and IR data and summary of mineral phases identified points to
some very relevant features about assemblages present in the preexplosion control sample (CS) and post-explosion samples and will be
discussed at time of presentation.
B115 Leuco Dye Detection of Latent Blood:
New Fluorescein Chemistry and
Other Promising Chromophores
Gabor Patonay, PhD*, Lucjan Strekowski, and Maryam Hojjat,
Georgia State University, Department of Chemistry, University Plaza,
Atlanta, GA 30303; James J. Krutak and Jeffrey Leggit, Federal
Bureau of Investigation, FBI Academy, ERF Building 27958A,
Quantico, VA 22135; Heather Seubert and Rhonda Craig, Federal
Bureau of Investigation, 935 Pennsylvania Avenue, NW, Washington,
DC 20535-0001
use this new latent blood stain detection kit, and how this kit is superior
to other techniques.
This presentation will describe new chemistry that significantly
improves latent blood detection, greatly increases the time window for
evidence collection and is easy to use. Additional leuco dye chemistries
can be tailored to avoid interferences. This new fluorescein chemistry
works even through paint.
Leuco dyes typically exhibit zero or very little fluorescence when
applied to surfaces. If latent blood residues are present leuco dyes are
oxidized and become fluorescent dyes. Fluorescin has been used frequently for this purpose. The chemistry used in the past has several disadvantages. The most significant of these is the rapidly diminishing contrast after application. Typically the investigator has a very limited time
to secure pictures of the site. After a few minutes fluorescin background
becomes fluorescent even where there is no blood residue thus effectively preventing photography of the crime scene. In addition to the
background difficulties, past chemistries need to be prepared at the site.
This new fluorescein chemistry works even through paint.
This paper discusses a new chemistry for fluorescin latent blood
detection. This new chemistry results in excellent contrast for several
hours and in a significant number of cases the contrast remains for days
or weeks. The FBI’s Evidence Response Team and DNA I unit worked
together with Georgia State University to validate this new fluorescin
chemistry for use in the field. This new chemistry requires no preparation at the crime scene. The kit is supplied in a ready-to-go format, i.e.,
the formulation is packaged as used. This ready-to-use format does not
adversely affect shelf life and it can be stored up to a year.
Several examples will be presented to illustrate how time and conditions affect contrast and false positive results. The performance of the
new fluorescin chemistry on different substrates will be discussed.
While this new chemistry is fully compatible with presently used light
sources, new truly portable light sources can be used as well. The use of
blue LED light sources will be discussed. These light sources are now
commercially available from several sources or can be made easily using
off the shelf components.
In addition to the visualization of latent bloodstains, this kit is also
suitable to observe latent fingerprints on certain surfaces. This feature
does not interfere with latent bloodstain detection due to the time delay.
In addition to the practical applications of this new kit, the presentation
will discuss the chemistry behind the kit, e.g., what components are
present in latent blood or fingerprint that would facilitate fluorescin oxidation. In addition to fluorescein, rhodamines can be prepared in their
leuco form and are oxidized by latent blood stains. This new rhodamine
chemistry requires no preparation at the crime scene similar to fluorescein. The longer wavelength absorption and fluorescence properties
of rhodamine can be very useful on certain substrates of high interference. Finally the paper will discuss typical interferences and how
those can be minimized.
Latent Blood, Leuco Dyes, Fluorescein
Mineralogical, Explosives, Criminalistics
80
* Presenting Author
B116 Dr. Edmond Locard and Trace Evidence
Analysis in Criminalistics in the Early
1900s: How Forensic Sciences Revolve
Around Trace Evidence
analyze certain trace evidence with a GC-MS or an FTIR just because
there is one available in the laboratory, without really thinking of the
pertinence of the characteristics analyzed. This can be avoided by asking
the right questions first.
Eric Stauffer, MS*, Applied Technical Services, 1190 Atlanta Industrial
Drive, Marietta, GA 30066
B117 Could Have, Would Have, Should Have:
The Utility of Trace Evidence
proper cognitive process that are applied to trace evidence analysis, that
forensic sciences revolve around the trace evidence itself and not the
analytical instruments used to analyzed it, and finally, some of Dr.
Locard’s work.
This presentation will impact the forensic community by demonstrating that forensic scientists will be able to apply the proper cognitive
process to the approach to trace evidence analysis, and that forensic sciences revolve around the trace evidence itself.
This presentation will introduce the original criminalistics
developed by Dr. Edmond Locard in the early 1900s. The writings of Dr.
Locard were revolutionary, and they constitute the strong base from
which modern forensic sciences evolved through science and technological advances. In the midst of this evolution, however, it is sometimes
forgotten that trace evidence is the real subject of interest in forensic sciences.
Today, it seems that a crime laboratory cannot perform any work
without the use of a gas chromatograph-mass spectrometer (GC-MS), a
Fourier Transform Infrared Spectrometer (FTIR), and Capillary
Electrophoresis (CE). While this holds true for some particular aspects
of forensic sciences, the absence of such instruments does not prevent
the criminalist from, first, properly collecting trace evidence at crime
scenes and, second, analyzing a great number of traces.
Dr. Locard was the pioneer of modern criminalistics, though he did
not have the modern instrumentation that is available in crime laboratories in the twenty-first century. Nevertheless, his work was not necessarily less reliable than that done today because he was cognizant of the
significance of his work and its limitations. However, he describes many
different traces that were observed and collected from crime scenes such
as prints from fingers, nails, lips, faces, feet, and teeth. He also describes
the observations of stains, such as grease, candle, mucus, feces, blood,
semen, rust, food, etc. The diversity of his observations is incredible and
it encompasses many potential traces that are not necessarily taken into
account today.
Forensic sciences should not revolve around the instrumentation
used to perform trace analysis, but around the trace evidence itself. This
is a very important concept that is emphasized throughout Dr. Locard’s
writings that seems to be sometimes disregarded in our modern times.
Dr. Locard’s method of reasoning is probably the most important
concept in forensic sciences, and the general process of trace evidence
analysis should include the following questions in this order:
• What is the trace?
• What is its composition and characteristics?
• How can this trace be linked to its origin or to another trace (or
what are the pertinent characteristics that will be used for the
comparison process)?
• How can that be accomplished (or what instrumentation/
technique is needed to analyze these characteristics)?
This cognitive process emphasizes the trace evidence and not the
techniques used to analyze it. It ensures that the criminalist understands
the importance of what is analyzed and, therefore, it establishes a proper
base for the interpretation of the results. Without having a clue about the
significance of the characteristics analyzed the forensic scientist cannot
properly interpret the results of the analyses, and therefore cannot reach
accurate conclusions. The development of many new analytical techniques seems to have led scientists away from the significance of the
trace evidence itself. It is not uncommon to see scientists trying to
* Presenting Author
Dr. Edmond Locard, Criminalistics, Trace Evidence Analysis
Max M. Houck, MA*, West Virginia University, 886 Chestnut Ridge
Road, Suite 309, Morgantown, WV 26506-6216; Sarah E Walbridge,
BS, Microtrace, Inc., 1750 Grandstand Place, Elgin, IL 60123
After attending this presentation, attendees will understand the time
and money-saving aspects of trace evidence examinations and why laboratories should retain trace evidence in their services portfolio
This presentation will encourage a greater awareness of the utility,
significance, and benefits of trace evidence analysis
Non-DNA forensic disciplines, particularly trace evidence, have
suffered from the persistent perception of their lack of scientific rigor.
Traditionally dismissed as “could have” evidence, trace evidence examiners have generally lacked the resources that have become common to
their DNA-oriented colleagues. This has lead to a general weakening of
the perception of the utility of trace evidence—even by trace evidence
analysts themselves. A weak perception, the authors feel, leads to a weak
analysis and a weaker interpretation.
Highlighting the most commonly voiced arguments against trace
evidence, this paper will present counter-arguments, backed with model
data, which emphasize the utility, and even the necessity, of trace evidence as a integral part of any forensic laboratory. Actual cases where
no DNA was examined, the DNA was examined but was not suitable for
analysis, or where a positive finding of DNA would not move the investigation forward (spousal sexual assault, for example) will be the most
obvious starting point for this presentation. Additional cases that demonstrate the potential specificity and strength of trace evidence analysis
will be discussed. Management issues will also be discussed showing
how trace evidence can save a laboratory time and money.
For example, in three hypothetical cases involving hairs in three
laboratories with differing sampling protocols, the laboratory employing
microscopical hair examinations has a more efficient and cost-effective
supply chain than laboratories that do not. Therefore, while hair examiners may be considered to be a “waste of time” by some laboratory
managers, they, in fact, can save both time and money.
Other actual and hypothetical examples will be discussed.
Lab
#1
($1500 per sample Analyzes DNA on
for mtDNA)
all Q hairs, plus
all K samples
Cost
Accuracy
Lab
#2
Only analyzes 2
hairs @ from
victim’s underwear and SAK,
plus all K samples
Lab
#3
Only analyzes 1
hair after microscopical exam from
each item, plus 2
K samples
Cost
Cost
Accuracy
Accuracy *
Small Case
2 positive/5 total
2 Known samples
$10,500 5 in 5
$9,000
4 in 5
$6,000
9 in 10
Medium Case
5 positive/15 total
4 Known samples
$36,000 20 in 20
$12,000
4 in 20
$9,000
9 in 10
Large Case15
positive/50 total
6 Known samples
$84,000 50 in 50
$15,000
4 in 50
$12,000 9 in 10
* Based on results published in Houck, M.M. and Budowle, B. JFS, V47, N5, 2002.
Trace Evidence, DNA, Management
81
B118 Future Crime Labs Without Trace
Evidence—Dysfunctional Dystopias
B119 Daubert Factors Applied to the
Examination of Footwear Evidence
Sandra Wiersema, BS*, FBI Laboratory, 2501 Investigation Parkway,
Quantico, VA 22135
awareness of the nature and importance of trace evidence.
The term trace evidence doesn’t refer to a single technique or technology. It is a misunderstood term.Trace evidence is an approach to
problem solving as much as it is an evidence category. The approach has
broad applicability to an extraordinary range of problems that arise in
criminalistics. This fact is not appreciated by people whose knowledge
of forensic science is derived from portrayals in the media or those of the
entertainment industry. Surprisingly, to some extent this is true of people
employed in forensic science. Trace evidence sections in some laboratories have been downsized as more resources are being invested in
DNA typing technology. It is easy for layman and scientist alike to
become enamored of high tech tools. We need to remind ourselves that
tools are rarely the solution. They need to be applied knowledgeably and
intelligently. Carefully framed scientific questions need to be posed
before the tools can be applied with maximum effect. Hopefully, predictions of the demise of trace evidence are premature. Trace evidence is an
essential contributor to case solutions in criminalistics and cannot be
allowed to wither. Trace evidence is inextricably intertwined with the
essence of criminalistics. It is not much of an exaggeration to assert that
“trace evidence is criminalistics.” Trace evidence approaches and assessments may define scientific problems in ways that allow the meaningful
application of techniques or tools such as elemental analysis or DNA
typing.
Tasks in criminalistics cannot be subdivided into a finite array of
predefined problems. Every case is different. The approach cannot be
selected from a reference book of methods. It cannot be known a priori.
Trace evidence approaches to the solution of casework problems
are essential to the future of criminalistics. If we lose sight of this point,
criminalistics could evolve to the extent where it is eviscerated of its
essence. It would be unrecognizable. It would cease to be science. In this
“Brave New World” crime laboratories would be composed solely of
technicians, instruments, and quality control managers. Investigators
would bring in evidence and select from a brightly colored McDonald’stype menu of available analyses. In this paint-by-the-numbers approach,
the investigator would be making physical evidence assessments of sorts
and the laboratory workers, I am reluctant to call them criminalists,
would carry out the prescribed analyses. The laboratory workers would
be mere technicians while the investigators, irrespective of their academic qualifications would be cast in the role of scientists. Some benefits might accrue. The item throughput might increase, and the resulting
error rates might be vanishingly small, but much of the work would be
of marginal value, or even irrelevant, and case solutions would suffer.
The resulting lab system might be good at doing the job right but it
would not be doing the right job. Sadly, perhaps this dystopia is closer
than we think. Action must be taken to reverse the drift in this direction.
The science in criminalistics is not embodied in carrying out tests and
operating computerized instruments. The science is in the all-important
front end assessment of the physical evidence that drives the selection of
the analyses to be performed and in the critical back end integration and
interpretation of the results of these tests that gives them meaning.
The purpose of this presentation is to acquaint the footwear
examiner with the Daubert/Kumho criteria and to offer suggestions for
answering these criteria as they relate to footwear evidence.
This presentation will provide answers to these criteria that the
author has used in a previous Daubert hearing along with suggestions
that can be used to demonstrate the way that comparisons are done and
the reliability of the methods used.
In Daubert, the United States Supreme Court has created a gatekeeping role for trial judges as to the admissibility of scientific expert
testimony. The Supreme Court has envisioned that trial courts would
conduct a factor-based analysis when determining whether the testimony
is reliable. The issues to be considered by the trial judge are:
1. whether the theory or technique can be and has been tested.
2. the existence and maintenance of standards controlling the
technique’s operation;
3. whether the theory or technique has been subjected to peer
review and publication;
4. the known or potential rate of error; and
5. general acceptance
Trace Evidence, Criminalistics, Dysfunctional Dystopias
82
The opinion noted that the factors did not comprise a definitive
checklist or test.
Initially, Daubert was applied only to “scientific” expert testimony.
In Kumho tire, the courts acknowledged that trial judges also have to
apply the Daubert analysis in the context of experience-based expert testimony. This means whether your discipline is considered to be “scientific” or “technical,” Daubert still applies.
This presentation will provide answers to these criteria that the
author has used in a previous Daubert hearing along with suggestions
that can be used to demonstrate the way that comparisons are done and
the reliability of the methods used.
The prosecutor or judge may suggest that the jury be allowed to
make the comparison between the footwear impression and the shoes in
question. The author will recommend examples that you can use which
should convince the court that this is not a good idea.
Avenues for further research and previous Daubert decisions will
also be discussed.
Daubert, Footwear, Evidence
B120 Analyzing Bar Soaps by Utilizing a Variety
of Optical and Chemical Techniques
Gerald M. LaPorte, BSc, MSFS*, Jeffrey Wilson, BS, MSF, Douglas
Shaffer, MS, and Robert Ramotowski, MS, United States Secret Service,
Forensic Services Division, 950 H Street NW, Washington, DC 20223
After attending this presentation, attendees will understand
methods to forensically examine and differentiate bar soap.
This presentation will present the forensic community with techniques to forensically identify and characterize bar soaps. This topic has
not been discussed (or infrequently mentioned) in the forensic literature.
As well, the authors will have a database of information which can be
made available to other analysts.
As a result of the anthrax poisonings that took place in October,
2001, and highly publicized news releases pertaining to chemical and
biological threats, law enforcement agencies have received numerous
hoaxes involving unknown material. Materials of unknown origin are
often submitted for forensic examination and can be analyzed using a
* Presenting Author
variety of analytical techniques. Recent evidentiary submissions and
forensic requests at the United States Secret Service have included
unidentified substances suspected to be soap. The perpetrators of these
crimes were obviously attempting to instill fear into the recipient by
making the victim believe they had been exposed to a chemical or biological agent. Investigators have requested identifications of the
unknown substances and comparisons with known material seized from
the suspect(s). After reviewing the forensic literature, there was very
little information specifically addressing the forensic examination of
soap bars. Therefore, the authors examined a variety of commercially
available soap bars to determine if the soaps could be characterized
using some of the common analytical equipment found in forensic laboratories. Fifty bars of soap were obtained and analyzed using optical
techniques, gas chromatography/mass spectrometry, thin layer chromatography, fourier transform infrared spectrophotometery, ultraviolet
and fluorescence spectrophotometry, and scannining electron
microscopy - energy dispersive spectroscopy. Numerous differences
between the soap specimens were identified and classified into a flow
chart/database so that other analysts can use the information in cases
involving the forensic analysis of suspected soap.
desorption/ionization (MALDI), Fourier transform infrared spectroscopy (FT-IR), electrospray ionization mass spectrometry (ESI-MS),
nanoelectrospray ionization mass spectrometry (nanoESI-MS) and
liquid chromatography/electrospray ionization mass spectrometry
(LC-MS).
All of these methods have shortcomings; several are
non-specific, some are not suitable for nonoxynol-9 in mixtures and
most involve techniques that are not readily available in most forensic
laboratories.
In this presentation, we will demonstrate a reproducible method for
detecting nonoxynol-9 using pyrolysis gas chromatography (PGC), an
instrument found in most forensic laboratories. Coupled with a mass
spectrometer (PGC-MS), examination and identification of nonoxynol-9
pyrolysis products in spermicidal lubricants is possible.
Pyrolysis and GC-MS conditions were varied using standard nonoxynol9 to optimize the method. Extraction methods and sample requirements
will also be discussed.
Soap Analysis, Chemical and Biological Threats, Bar Soap
B122 Forensic Analysis of Architectural
Paints Utilizing Raman Spectroscopy
B121 Analysis of Nonoxynol-9 in Condom
Lubricants via Pyrolysis Gas
Chromatography-Mass
Spectrometry (PGC-MS)
Philip R. Antoci, MS*, W. Mark Dale, BA, New York City Police
Laboratory, 150-14 Jamaica Avenue, Jamaica, NY 11432; Peter R.
DeForest, DCrim, John Jay College of Criminal Justice, New York, NY
10019; John A. Reffner, PhD, SensIR Technologies, Danbury, CT
06810; William Grossman, PhD, Hunter College, New York, NY
10021; David C. Locke, PhD, Queens College, Flushing, NY 11367
Christopher R. Bommarito, MS, Michigan State Police Forensic
Science Division, 7320 North Canal Road, Lansing, MI 48913; Elaine
Dougherty, BS*, School of Criminal Justice, Michigan State University,
560 Baker Hall, East Lansing, MI 48823
After attending this presentation, attendees will have been introduced to Raman Spectroscopy as an analytical technique that can aid in
the forensic analysis and comparison of architectural paints.
The use and advantages of Raman spectroscopy in the forensic
analysis of architectural paints and pigments will be presented. In this
study, samples of architectural paint pigments and pigmented architectural paints were analyzed by Raman spectroscopy. The data presented
will show that Raman spectroscopy can aid the forensic scientist in the
discrimination of architectural paints. This research documents a new
approach to the analysis and characterization of architectural paints.
Although paint evidence is common trace evidence, the forensic analysis
of architectural paints has not received significant attention in recent
years.
The forensic scientist must not be concerned only with positive
associations of physical evidence. As an unbiased finder of truth, the
forensic scientist must be cognizant of analytical methods that provide
the best discrimination among similar samples. Forensic scientists must
consider all tests and analytical procedures that may result in the discrimination between paint samples. The greater the number of analytical
techniques and methods the forensic scientist has available and utilizes
to analyze physical evidence, the greater the probability of discriminating between items of evidence. The ability to provide greater discrimination among similar paints will result in a reduction of false inclusions. An analysis employing a battery of discriminating techniques that
shows consistency between samples will result in a stronger association.
Forensic paint evidence is received at the forensic laboratory in the
course of many types of criminal investigations. Such investigations
include but are not limited to: forced entries, burglaries, robberies,
assaults, vehicular hit and runs, abductions, rapes, and homicides. The
transfer of paint occurs from miscellaneous contacts of painted objects
with each other or contacts of painted surfaces with non-painted surfaces. Mutual transfers may not always be equal in quantity or quality.
In some cases, very light or minimal transfers can occur, these types of
transfers often present a challenge to the forensic scientist. Minimal
transfers can make the characterization, identification, and association of
After attending this presentation, attendees will understand the use
of pyrolysis gas chromaography mass spectrometry for presence of
condom lubricants in sexual assault cases.
This presentation will demonstrate new techniques for detection of
condom lubricants.
Many sexual offenders are now utilizing condoms in sexual
assaults. Because of this fact, forensic examiners increasingly need to
examine swabs for condom lubricants in criminal sexual conduct cases
in which no sperm is detected.
Examination of condom lubricants is problematic due to both contamination issues and the nature of the chemical constituents of the lubricants. Due to the presence of bodily fluids and other lubricants commonly present on vaginal swabs it is preferable to utilize a chemical separation technique such as gas (GC) or liquid chromatography (LC) in
examination of the swabs. The high molecular weight and chemical
nature of the surfactants, however, make introduction and examination
of samples by these instruments problematic.
One of the most common lubricant components, the spermicide
nonoxynol-9 is a harsh detergent composed of a nonylphenol with a
polyoxyethylene chain of varying lengths. It has a range of molecular
weights differing by 44 amu, the weight of one ethylene oxide
(-OCH2CH2-) group. The analysis of this detergent by traditional
electron impact (EI) ionization GC-Mass Spectrometry is not useful. It
is an extremely viscous liquid that is not volatile enough get through the
injector and into the column. Its large molecular mass (average 616 amu)
and both hydrophilic and hydrophobic ends also contribute to difficulties
in analysis by GC-MS.
Several other methods for detection of nonoxynol-9 have been
explored, including thin layer chromatography (TLC), desorption
chemical ionization mass spectrometry (DCI-MS), matrix-assisted laser
* Presenting Author
Pyrolysis, Condom, Nonoxynol-9
83
items of evidentiary value very difficult. Raman spectroscopy can be
applied to the analysis of very small samples; and therefore is ideal for
the analysis of this type forensic sample.
Over the last five years, the technology for collection of Raman
spectra has advanced, transforming Raman spectroscopy from a research
curiosity into a practical analytical methodology. Raman spectroscopy
holds great potential for analyzing all types of forensic evidence.
Advances in Raman instrumentation and technologies, have resulted in
instruments that are far different from their predecessors. The cost of
these new commercial instruments has decreased sufficiently that
forensic laboratories can now afford them.
Prior to application, architectural paint is a liquid medium containing a mixture of a solvent(s), dispersant(s), binder(s), pigment(s),
and additive(s). When paint is applied to a surface it dries to form a thin
film or coating that contains all but the solvent portion of the mixture.
Architectural coatings are applied to both the interior and exterior surfaces of building structures. Over 600 million gallons of architectural
paint are applied to the interior and exterior surfaces of building structures each year.
Paint can contain both organic and inorganic materials. The fact
that paint binders are poor Raman scatterers is advantageous. A large
number of pigments used in architectural paints are strong Raman scatterers. This phenomenon allows for the identification of pigments in
architectural paints. In addition, with the requirement of little or no
sample preparation coupled with the speed at which Raman spectral data
can be acquired, the discrimination of architectural paints can be accomplished in a matter of minutes.
Multiple applications of paint result in the production of a series of
layers. Samples consisting of single layer coatings of paint as well as
intact, multiple-layered paint chips were analyzed using Raman spectroscopy. A semi-quantitative analysis of the Raman spectral data
obtained resulted in the discrimination of similar white paint samples.
The analytical data presented will support the utility and value of Raman
spectroscopy in the analysis architectural paints.
Raman, Paint, Pigments
B123 Near Infrared Microspectral
Analysis of Trace Evidence
Jumi Lee, PhD*, CRAIC Technologies, 2400 North Lincoln Avenue,
Altadena, CA 91001
After attending this presentation, attendees will understand techniques for analyzing representative trace evidence samples in the near
infrared. Results from the analysis of basic types of trace evidence
useful for aiding scientists with their own identifications.
This presentation will demonstrate the analysis of representative
types of trace evidence in a spectral region that has not been previously
explored.
The examination of trace evidence by UV-visible microspectroscopy has been an integral part of trace evidence examination for
thirty years. Many types of fibers, paints and other evidence of concern
have been analyzed in detail for both their color and UV characteristics.
However, almost no work has been done investigating the near infrared
spectral characteristics of such samples and determining the utility of
microspectroscopy in this region as a comparative technique.
By the near infrared region, we mean the spectral range from 700
nm to 2100 nm. This work shows the results of microspectral analysis
of a number of representative samples in the spectral region from 700 to
2100 nm as well as an analysis of the results. The major features are
described for use by other examiners in their casework.It also discusses
the best techniques for preparing samples for analysis in the NIR region.
Near Infrared, Trace Evidence, Microspectroscopy
84
B124 Identification of Odor Signature Chemicals
Used by Detector Dogs to Locate Drugs
and Items Containing Drug Odors
Including Currency
Kenneth G. Furton, PhD*, Norma Lorenzo, MS, Brad Young, BS, Ya-Li
Hsu, MS, and Stefan Rose, MD, International Forensic Research
Institute, Department of Chemistry and Biochemistry, University Park,
Florida International University, Miami, FL 33199
This paper describes the identification and quantification of odor
signatures used by detector dogs to locate controlled substances and the
significance of these finding in confirming dog alerts to items associated
with controlled substances including currency
This presentation will demonstrate that law enforcement detector
dogs are trained to alert to significant quantities of unique odor chemicals associated with controlled substances and not found in common
over the counter preparations or circulated currency thus strengthening
the value of dog alerts to items as part of an investigation.
This paper describes the use of headspace solid-phase microextraction (SPME) combined with gas chromatography mass spectrometry
(GC/MS) to identify the signature odors that law enforcement certified
detector dogs alert to when searching for drugs. Background information
is provided on the many types of detector dogs available and specific
samples highlighted in this paper are the drugs cocaine and 3,4Methylenedioxy-N-Methylamphetamine (MDMA or Ecstasy). Studies
include the analysis and identification of the headspace (fingerprint) of
a variety of samples, followed by completion of double-blind dog trials
of the individual components in an attempt to isolate and understand the
target compounds that dogs alert to. SPME/GC/MS has been demonstrated to have a unique capability for the extraction of volatiles from the
headspace of forensic specimens and shows great potential to aid in the
investigation and understanding of the complicated process of canine
odor detection. Major variables evaluated for the headspace SPME
included fiber chemistry and a variety of sampling times ranging from
several hours to several seconds and the resultant effect on ratios of isolated volatile components. CW/DVB and PDMS SPME fibers proved to
be the optimal fiber types. Field studies with detector dogs have demonstrated possible candidates for new pseudo scents as well as the potential
use of controlled permeation devices as non-hazardous training aids providing consistent permeation of target odors.
The results demonstrate that SPME/GC/MS combined with field
tests using certified detector dogs is an effective method for identifying
active odor signature chemicals in forensic specimens. For the drug
studies it was found that passive adsorption using DFLEX devices containing activated charcoal was not sensitive enough to recover signature
odors from the headspace of MDMA tablets. Only when very large
samples (i.e., greater than 250 grams) were extracted for extended
periods of time (i.e., 1 week) were odors reliably detected. However,
with headspace SPME it was found that with the implemented use of the
CW/DVB and PDMS fibers with 3 hour extraction times it was possible
to obtain consistent signature odors from the headspace of a single
MDMA tablet. Many compounds of interest were found to be present in
the headspace composition of the MDMA tablets tested, including
piperonal, MD-P2P and methamphetamine. Through examinations of
different tablets, however, it was concluded that the metamphetamine
found within certain tablets were present due to its addition as an
impurity or adulterant and not the direct result of synthetic manufacturing and that piperonal and MD-P2P were the common chemicals seen
in all samples tested.
In studies where different over the counter tablets were analyzed, it
was concluded that none of the headspace compounds found within
these tablets were present in the headspace of MDMA tablets, therefore
making the possibility of false positive alerts from the canines in association with these commonly encountered tablets unlikely. Field studies
* Presenting Author
directly focusing on the signature odor of MDMA have shown that
canines are alerting to approximately 10 ¡V 100 mg of the piperonal
compound that is found exclusively in MDMA tablets. Since MDMA
manufactured through different synthetic routes can yield different signature chemicals, it is important to perform ongoing studies of headspace odors from current street samples and more than one MDMA
training aid may be required for optimal performance in the future.
The dominant cocaine odor chemical has been confirmed to be
methyl benzoate via spiked samples as well as controlled delivery
devices with threshold levels of 1-10ng spiked methyl benzoate or 0.1-1
ng/sec odor diffusion. The evaporation rate of methyl benzoate from circulated and uncirculated U.S. currency has also been studied in detail.
The amount of methyl benzoate on currency decreased exponentially.
Evaporation rates varied considerably (2 to 2000 ng/sec) depending on
conditions, decreasing with increasing number of bills and the covering
of the currency. The levels of signature odor chemicals needed to initiate
consistent alerts from law enforcement detector dogs and the lack of significant levels of these chemicals in common over the counter items or
circulated currency enhances the significance of dog alerts to items as
part of an investigation.
Drugs, Detector Dogs, Currency
B125 The Effects of Adsorption-Based
Extraction Methods on the Recovery
of Aliphatic and Aromatic Compounds
in Medium Petroleum Distillates
Julia A. Dolan, MS*, Bureau of Alcohol, Tobacco, Firearms &
Explosives, Forensic Science Laboratory, 6000 Ammendale Road,
Ammendale, MD 20705; Eric Stauffer, MS, Applied Technical
Services, Inc., 1190 Atlanta Industrial Drive, Marietta, GA 30066
Following this presentation, the attendee will have a greater understanding of the following: (1) the importance of relative aliphatic:aromatic concentrations in classifying medium-range petroleum products;
(2) how adsorption-based extraction methods can affect these ratios with
respect to the recovery of a medium petroleum distillate from simulated
fire debris; and (3) the effects of ignitable liquid concentration on the
aliphatic:aromatic ratios.
This presentation will assist examiners in understanding the phenomena associated with the extraction process and how it may skew
data. This knowledge may ultimately assist in minimizing the likliehood
of false inclusions or exclusions when comparing data.
Classification of ignitable liquids in accordance with voluntary
consensus-based standards published by the American Society of Testing
and Materials has become increasingly specific, relying upon both the
chemical composition and the boiling point range of submitted ignitable
liquids. Specifically, the classification of petroleum distillates and
dearomatized petroleum distillates has been distinguished, such that
each represents a distinct class.
In the first part of this study presented in 2003, the compositions of
a variety of medium-range ignitable liquids were examined with regard
to their relative proportions of aromatic and aliphatic content, as represented through the use of extracted ion profiles (EIPs). A variety of
commercially available products in the medium range exhibit a broad
range of compositions with respect to the proportion of aromatic compounds relative to the major aliphatic compounds present. It was shown
that there are not separate and distinct ranges of aromatic content for
dearomatized products, distillates, and blends; rather, there is a continuum.
This portion of the study examines how the extraction process
affects the relative proportion of aliphatic and aromatic compounds
recovered. Samples of simple and complex substrates were spiked with
varying volumes of a medium petroleum distillate. These samples were
* Presenting Author
then extracted in accordance with ASTM 1412-00 Standard Practice for
Separation of Ignitable Liquid Residues from Fire Debris Samples by
Passive Headspace Concentration With Activated Charcoal. Data was
then examined to determine if the process of extracting the liquid from
debris affected the relative proportion of aromatic and aliphatic compounds. Using data from the neat liquid for comparison, data from
extracted samples were compared to note the effects of substrate type
and volume of liquid. Examination of the data revealed that the primary
factor affecting the recovery of aromatic and aliphatic compounds relative to one another was the concentration of ignitable liquid in the
sample. A greater volume of medium petroleum distillate on the sample
resulted in a greater recovery of aromatic compounds relative to
aliphatic compounds. Also affecting the recovery and subsequent ratios
was the nature of the debris. The more complex charred debris samples
showed a greater relative recovery of aliphatic compounds compared
with their recovery from a simple, non-charred substrate. These results
are consistent with the hypothesis that competitive adsorption can alter
aliphatic:aromatic ratios due to the fact that charred debris can provide
sites for adsorption. This also further demonstrates that overloading
adsorption sites can lead to skewing of data not only relative to boiling
point as previously reported,1 but also with regard to relative
aliphatic:aromatic ratios. These factors studied which were found to
affect recovery—amount of ignitable liquid present, and nature of
sample substrate—are beyond the control of the laboratory analyst in
actual casework. It is therefore incumbent upon the analyst to be aware
of the significant effects of the extraction process when utilizing
aliphatic:aromatic ratios in the classification process or when comparing
data from extracted samples with one another or to data from a liquid
source.
1 Newman, RT, Dietz, WR and Lothridge, K. “The Use of Activated
Charcoal Strips for Fire Debris Extractions by Passive Diffusion. Part I:
The Effects of Time, Temperature, Strips Size, and Sample
Concentration,” (1996) Journal of Forensic Science, Vol. 41, No. 3, 361370.
Fire Debris Analysis, Extraction, Aromatic Content
B126 Detailed Chemical Composition of
Ignitable Liquids Determined With
Comprehensive Two-Dimensional Gas
Chromatography-Mass Spectrometry
Glenn S. Frysinger, PhD*, U.S. Coast Guard Academy, Department of
Science, 27 Mohegan Avenue, New London, CT 06320-8101
comprehensive two-dimensional gas chromatography with mass spectrometry detection produces detailed chemical composition information
about ignitable liquids that can be used to identify these substances in
complex fire debris samples.
The presentation will describe advances in analytical chemistry that
will have a significant impact on the field of forensic fire debris analysis.
Comprehensive two-dimensional gas chromatography methods provide
an order of magnitude improvement in the chemical separation of
complex mixtures that will lead to enhanced identification of ignitable
liquids.
Comprehensive two-dimensional gas chromatography with mass
spectrometry detection (GCxGC/MS) is a three-dimensional hyphenated
analytical method that is ideal for exploring the complex chemical composition of ignitable liquids. The increased separation and identification
capability of GCxGC/MS represents a significant analytical advantage
over traditional gas chromatography (GC) and gas chromatographymass spectrometry (GC/MS) methods for the analysis of complex mixtures. GCxGC uses two chromatography columns with different stationary phases that are coupled serially by a modulator. Ignitable liquid
85
compounds can be separated by a combination of volatility and polarity
or shape mechanisms to produce a two-dimensional chromatogram with
hundreds of compound peaks distributed across a two-dimensional
retention time plane. The two-dimensional separation is a significant
improvement over traditional GC separations because many more peaks
are resolved and the peaks are arranged into groups according to their
chemical class. In this way, alkanes, alkenes, cycloalkanes, aromatics,
and oxygenated compounds are rapidly classified. When GCxGC separations are coupled with a mass spectrometer, each two-dimensional
chromatogram peak has an associated mass spectrum that leads to
accurate identification with mass spectral libraries.
In this work, the chemical composition of common ignitable liquids
such as paint thinner, lacquer thinner, gasoline, and kerosene will be
examined. Compounds separated by GCxGC will be assigned to a
chemical class and identified by mass spectrometry. The comprehensive
chemical information will be used to fingerprint ignitable liquids in the
presence of chemical interferences from fire debris combustion and
pyrolysis products.
Arson, Gas Chromatography-Mass Spectrometry, Ignitable
Liquids
B127 Evaluation of the Effectiveness of Nylon
Bags as Packaging for Fire Debris
Aleksandra E. Stryjnik, BSc* and Robert Hong-You, BSc, Centre of
Forensic Sciences, 25 Grosvenor Street, Toronto, Ontario M7A 2G8,
Canada
After attending this presentation, attendees will have been presented with the limitations associated with the use of nylon bags as packaging for fire debris samples with respect to the possible loss or crosstransfer of ignitable liquid vapors.
This study confirmed that nylon bags are suitable packaging for fire
debris samples if properly sealed. Heat-sealing, although the most
effective at retaining ignitable liquid vapors, is not always practical in
the field and is not easily reproducible. Folding the opening of the nylon
bag three times and then duct taping the fold is a satisfactory alternative
that limits the loss of ignitable liquid vapors. The swan neck seal proved
to be least effective, however, this study shows that no cross transfer
occurred between nylon bags sealed in this way when stored for an
extended period of time. Nonetheless, efforts to minimize the possibility
of cross-contamination, such as separating samples from different
sources, are recommended.
A proper container for packaging of fire debris samples must be
effective at retaining ignitable liquid vapors. Glass Mason jars, metal
cans, and nylon bags are the accepted packaging. At this laboratory the
preferred packaging is glass Mason jars; nylon bags are recommended as
an alternative when the fire debris is too large to fit the preferred packaging. A study was undertaken to explore the effectiveness of Grand
River® nylon bags with a swan neck seal, as typically received at this
laboratory. The effectiveness of these bags, different sealing techniques,
and the possibility of cross-transfer of ignitable liquid vapors between
adjacent bags were evaluated. Analysis involved dynamic headspace
sampling using tubes packed with Tenax® adsorbent and gas chromatography with flame ionization detection (GC-FID) or gas chromatography-mass spectrometry (GC-MS).
An experiment was designed to evaluate the loss of several classes
of ignitable liquid vapors. A solvent mixture, gasoline, kerosene, and
light, medium and heavy petroleum distillates were used. Paper towels
spiked with 50µL of an ignitable liquid were packaged in nylon bags
with a swan neck seal and subsequently placed into a clean glass Mason
jar, which was immediately capped. Room temperature analysis of the
glass Mason jar headspace was conducted to determine whether any
vapors had escaped from the nylon bags. The results indicated loss of
ignitable liquid vapors regardless of product class. Further work was
86
undertaken to determine whether the loss occurred through the walls of
the nylon bags or through the seal.
The effectiveness of different sealing techniques was evaluated as
above using paper towels spiked with 50µL of gasoline. The sealing techniques tested were a swan neck seal, a triple fold sealed with duct tape, a
single heat seal and a double heat seal. Single and double heat seals were
the most effective at retaining ignitable liquid vapors, as gasoline was not
identified in the headspace of the glass Mason jars. Low levels of incomplete gasoline patterns, which would not meet this laboratory’s criteria for
identification of gasoline, were observed from nylon bags that were sealed
with duct tape. Identifiable gasoline patterns were seen in the results from
the nylon bags sealed with a swan neck seal.
The effect of double bagging with a swan neck seal was also investigated using gasoline. The results indicated no significant advantage,
with respect to retaining ignitable liquid vapors, over a single bag with
a swan neck seal.
The possibility of cross transfer of ignitable liquids between nylon
bags with a swan neck seal was explored. Two nylon bags were placed
together in a cardboard box: one nylon bag contained paper towels
spiked with 1mL of an ignitable liquid; and, the other contained clean
paper towels. As in the first part of the study, a solvent mixture,
gasoline, kerosene, and light, medium and heavy petroleum distillates
were used. The cardboard box was sealed with tape and stored for
approximately five months. No ignitable liquid vapors were identified
in the box or the adjacent nylon bag, despite the presence of visible
liquid or strong positive results in the spiked nylon bag.
This study confirmed that nylon bags are suitable packaging for fire
debris samples if properly sealed. Heat-sealing, although the most
effective at retaining ignitable liquid vapors, is not always practical in
the field and is not easily reproducible. Folding the opening of the nylon
bag three times and then duct taping the fold is a satisfactory alternative
that limits the loss of ignitable liquid vapors. The swan neck seal proved
to be least effective, however, this study shows that no cross transfer
occurred between nylon bags sealed in this way when stored for an
extended period of time. Nonetheless, efforts to minimize the possibility
of cross contamination, such as separating samples from different
sources, are recommended.
Fire Debris Analysis, Packaging, Nylon Bags
B128 Advances in the Smoke Odor
Analysis in Fire Investigations
Robert Large, PhD*, Chris A.J. Harbach, PhD, and Rachael
Stockford, MSc, M-Scan Ltd, 3 Millars Business Centre, Fishponds
Close, Wokingham, Berkshire RG41 2TZ, United Kimgdom; Mark
Rogers, PhD, M-Scan Inc, 606 Brandywine Parkway, West Chester,
PA 19380
After attending this presentation, attendees will have been presented with data illustrating how smoke odor analysis can be used in
arson and homicide investigations and to allow the delegate to develop
an appreciation of a new forensic tool.
This presentation introduce a new forensic tool.
Smoke odor comprises a variable range of individual volatile
organic compounds (VOCs), which can be present above or below published odor thresholds. The detailed distribution of VOCs in the vapor
phase depends upon the particular combination of materials being
pyrolysed in the fire in question, for example wood, polystyrene, polyolefins, polyurethanes, polyesters, PVC, nylons, methacrylates, tissue
and various accelerants. The VOC products in question can accumulate
efficiently on both adsorbent materials at the scene of the crime and on
the clothing and hair of a suspect exposed for a period to smoke from the
fire in question. They can then be detectable organoleptically by both
scene of crime and arresting officers.
* Presenting Author
The adsorbed pyrolysis products are amenable to established headspace analytical procedures, involving concentration from sealed nylon
pouches onto conditioned Tenax porous polymer and thermal desorption/GC-MS analysis. Approximate quantification allows comparison with an extensive database of odor thresholds. Pyrolysis-EIMS
analysis of materials from the scene of the crime can be used as a validation procedure to verify the probable source(s) of volatile products in
question.
The pyrolysis products are present in smoke at typically ppb concentrations. Much care needs therefore to be devoted to data interpretation and the selection and analysis of appropriate control samples.
Although the TD/GC-MS methodology is similar, this presents a significantly more challenging analytical requirement than typically encountered in the detection and characterization of accelerant residues on fire
debris samples.
The M-Scan smoke odor method will be described and illustrated
by reference to (a) the first English criminal case in which this approach
has been used to tentatively link a suspect to a particular fire, and (b) a
series of controlled combustion experiments involving known combinations of materials.
Pyrolysis, Smoke, Odor
B129 An Atypical Cause of Fire:
CCTV Evidence to the Rescue
Rebecca S. Pepler, BSc, MPhil*, Anglia Polytechnic University,
Departmentt of Forensic Science & Chemistry, East Road, Cambridge,
Cambridgeshire CB1 1PT, England
After attending this presentation, attendees will recognize the
importance of CCTV evidence when investigating a large fire scene;
understand how civil litigation can direct and/or curtail an investigation;
implement a multidisciplinary and multi-agency investigation; learn of
an unusual substrate for spontaneous combustion; and learn how to
confirm the occurrence of spontaneous combustion using laboratory
facilities.
This case study will highlight the differences between the UK and
the USA in the way that civil litigation can influence the extent of a fire
investigation. In addition, it will demonstrate the large differences in the
way in which fire investigation is undertaken in the UK and the USA.
The presentation will reinforce the basic methodology of fire investigation, refresh investigator’s memories concerning aspects that should
not be overlooked and how best to exploit a multi-agency approach.
Furthermore, the case will reveal an unusual substrate that is capable of
self-heating, thus making investigators aware of the potential of this
material to spontaneously combust. The presentation will also explain
how best to use the available laboratory facilities to confirm or refute
initial suspicions of self-heating.
On a dry October Sunday afternoon a small fire was discovered in
the external storage area of a large expanded foam factory. Due to a
number of factors, including a strong south easterly wind, the fire escalated into one of London’s largest fires for a decade and resulted in a £26
million loss. The ensuing investigation lasted several months and is an
excellent example of a multi-disciplinary and multi-agency approach.
The potential civil litigation meant that the investigation incorporated
many facets of fire investigation, from sprinkler systems to fire fighting
procedures, and highlights the (sometimes frustrating) role that civil litigation plays in directing an investigation.
CCTV evidence proved invaluable in identifying the point of origin
of the fire. Without the video evidence, the cause of the fire may well
have been recorded as deliberate ignition. As it turns out the fire was initiated in a much more unusual manner. Subsequent laboratory tests confirmed the initial conclusions and demonstrated that the fire was caused
by self-heating (spontaneous combustion).
Spontaneous, Combustion, CCTV
* Presenting Author
B130 Cooperation Amongst Professionals:
A Complex Case Study of a Suspicious
Fire in Southern China
Sheilah E. Hamilton, PhD*, University of Glamorgan, School of
Applied Sciences, Pontypridd, Wales CF37 1DL, United Kingdom;
Zennon C.P. Cheng, Zennon & Pierre Marine Services, Ltd., 5/F
Greatmany Centre, 109-115 Queen’s Road East, Hong Kong, SAR PRC
The goal of this presentation is to show how the successful investigation of an unusual and complex international fraud case, spanning
three different countries, relied on the input of professionals with greatly
differing skills and experience before the scam was confirmed.
This presentation will demonstrate how the same skills used by
forensic professionals in criminal cases can be used in the investigation
of civil enquiries. It also underlines the importance of co-operation
between different disciplines.
During the early morning of 7 February 1999 a burning container
vehicle was reported at the foot of a steep slope at the side of the main
road in a remote area between Panyu City and Shunde in Guandung
province, Southern China. The vehicle consisted of a tractor, trailer and
a container which was reputed to contain 7,500 state-of-the-art PC
mother boards which had originated in Taiwan and been transhipped to
Hong Kong before arriving on the Chinese mainland.
Initially the fire was attended by local Chinese fire and police
officers who took two photographs but were not qualified to investigate
the cause of the fire. The driver of the vehicle had disappeared and was
never located. Later the same day the tractor, trailer, container and ~25%
of the cargo of mother boards were removed to a nearby police pound
where they remained until the tractor, trailer and mother boards were
claimed on payment of a fine for damage to surrounding crops at the
location of the accident. The empty container was not claimed and
remained at the police pound.
On March 31, 1999, a change in survey personnel occurred and the
new investigation team included the current authors. Despite the passage
of time, when the scene of the fire was visited approximately 500 mother
boards remained strewn on the ground and these were collected for
further examination together with other potentially useful items such as
wrapping material.
The investigation team included a fire investigator/forensic scientist, a chartered surveyor and a computer expert. The different skills of
each member were needed to determine that this was an elaborate fraud
which, had the insurance claim been upheld, would have resulted in the
payment of a large amount of money.
This presentation will show the different reasons why the authors
concluded that (1) the fire was a deliberate act, and (2) the value of the
motherboards was a small fraction of that claimed.
Fraud, China, Cooperation
B131 The Business Case for Using Forensic DNA
to Solve and Prevent Sexual Assault Cases
Ray A. Wickenheiser, BSc*, Acadiana Criminalistics Laboratory, 5004
West Admiral Doyle Drive, New Iberia, LA 70560
The learning objective of this presentation is to acquaint the
forensic scientist and forensic science administrator with the concept
and practical application of cost benefit analysis to the area of forensic
science. By applying this analysis to the specific topic of “no suspect
sexual assault,” a business case for forensic science will be amply
demonstrated in a manner that all can relate to, dollars and sense.
Through application of a cost benefit analysis, and development of
a sound business case, the forensic community will better demonstrate
the value of its service. The application of this method to cases of “no
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suspect “sexual assault demonstrates a many fold return on investment.
Ultimately, demonstrating the dollar value of using forensics to solve
and prevent crime will lead to appropriate levels of funding to meet the
requirements of society.
The elements truth and justice are very difficult to quantify and
measure. While no one doubts the value of forensic science in the establishment of truth, and hence justice, through the objective means of
science, many government forensic labs are experiencing difficulty in
securing adequate financial resources to meet their goals. Only one
quality of work is possible with justice at stake, the highest. The consequence of error is grave, so quality cannot be sacrificed for the sake of
output. There are high requirements to provide this top quality.
Education, ongoing training, proficiency testing, documentation, and
accreditation itself, add to the cost of providing the forensic service
deserved by all members of society. More than ever, forensic laboratories have become the victims of their own success. The demand for
quality service has outstripped the financial resources available and
required to deliver the services needed.
With expenditures of all levels of government scrutinized more
than ever, levels of funding are very tight. Retaining funding levels is
difficult, let alone increasing funding to expand services to meet
increasing levels of demand, and the high cost of technology. Making a
business case for forensic science involves placing a dollar value on the
costs and the benefits of forensics. While it is inherently very difficult,
establishing dollar cost and benefits demonstrates the real value of
forensics as an investment. Furthermore, the value of the investment in
forensics can be compared to expenditures and benefits of other programs.
Use of the very specific forensic case type of “no suspect” sexual
assault provides the framework for this business case. Cases of sexual
assault where the perpetrator is unknown to the victim are seldom
solvable without the assistance of forensic science. With the help of
recently established DNA databases, a suspect can be developed, and
cases solved, which were previously unsolvable. There is a high level of
recidivism in the case of sexual assault, where the same offender
commits a number of assaults. Early detection and apprehension of
offenders not only solves outstanding cases, but prevents future assaults
as well. The cost of providing forensic DNA service can be quantified.
The benefits of applying forensics to solve cases of sexual assault can
also be calculated, as well as the value of preventing future sexual
assaults. In this manner, a cost benefit analysis can establish the high
value of forensic DNA in quantifiable terms. With this very high return
on investment, the case can be made much more easily for adequate
resources to meet the level of demand needed by society.
Prior to 1999, the examination of sexual assault kits was the responsibility of the NYPD lab.The lab was only performing serological testing
on those cases going to court, and as a result, the number of sexual
assault kits not being examined accumulated rapidly. In the year 2000,
CODIS was implemented. The issue arose as to how the NYPD was
going to handle the large backlog of cases that had accumulated. The
NYPD backlog project was developed to examine approximately 17,800
cases regardless of whether these cases had been previously examined.
Three independent forensic laboratories, were contracted to examine or
re-examine these cases and the Office of the Chief Medical Examiner
reviewed all of the data and entered it into CODIS.
On March 19, 1991, a seventeen-year-old female was sexually
assaulted and robbed on the rooftop of her building. As a result, a sexual
assault kit was taken but no DNA testing was conducted. The victim was
able to provide the police with a detailed physical description of her
assailant. The man was subsequently apprehended, placed on trial, and
convicted of rape, sodomy, and robbery. The defendant claimed his innocence throughout the trial.
In 2003, Orchid Cellmark received the sexual assault kit from the
aforementioned case. Their analysis found the presence of sperm on the
vaginal slide. A cutting of the vaginal swab was submitted for STR DNA
testing resulting in a male DNA profile. A report was generated by
Orchid Cellmark and all results were forwarded to the OCME. The
results were analyzed by the Department of Forensic Biology, where the
male DNA profile was entered into the database to be uploaded into
CODIS. The laboratory was subsequently notified that the profile
uploaded for this case resulted in a match with a convicted offender.
This investigative lead was analyzed and determined that the profile
from the convicted offender was the same as the DNA profile obtained
from the vaginal swab. However, this convicted offender was not the
same man that was previously convicted of the assault.
The Manhattan District Attorney’s office requested that the sexual
assault kit be re-examined by the Department of Forensic Biology. The
presence of semen on the vaginal swab was confirmed and a cutting was
submitted for DNA analysis. An oral swab was obtained from the victim
to ensure that the kit being examined was indeed from her. DNA typing
of the vaginal swab corroborated the results that were obtained by
Orchid Cellmark. The man that was serving time in prison for twelve
years had been wrongfully convicted. On May 19, 2003, the man serving
time for sexual assault was informed that all charges against him had
been dismissed and that he was now a free man. Unfortunately, due to
the statute of limitations regarding sexual assault, the convicted offender
found to match this case cannot be prosecuted.
NYPD, NYPD Backlog Project, CODIS
Business Case, DNA, Sexual Assault
B132 NYPD Backlog Project Exonerates
Man After 12 Years
Linda Frese, MS*, Mary Quigg, BS, Howard Baum, PhD, and Robert
Shaler, PhD, Office of the Chief Medical Examiner, Department of
Forensic Biology, 520 First Avenue, New York, NY 10016
After attending this presentation, attendees will have been presented with a case in which the NYPD backlog project, and CODIS,
resulted in the exoneration of a man falsely incarcerated for twelve
years.
This presentation displays how CODIS is such a powerful tool. Not
only can it be used to aid in prosecuting offenders but also help to exonerate those wrongfully convicted of a crime. We would not have been
able to exonerate the man in this case without the help of the CODIS
database.
This talk will discuss the history of the NYPD backlog project and
how the re-analysis of one case exonerated a man convicted of sexual
assault, and resulted in a DNA match with a convicted offender.
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B133 The Mother of All DNA Contracts - 16,000
Sexual Assault Kits and Counting (Part I)
Marie Samples, MS*, Robert C. Shaler, PhD, and Howard J. Baum,
PhD, New York City Office of Chief Medical Examiner, Department of
Forensic Biology, 520 First Avenue, New York, NY 10016; Karen
Dooling, MS, Nassau County Medical Examiner’s Office, 2251
Hempstead Turnpike, Building R, East Meadow, NY 11554
The attendee will learn the trials and tribulations associated with a
very large casework DNA contract.
Since outsourcing seems to be on the upswing in order to reduce
DNA analysis backlogs, presentation about the experiences of New York
City may help others in a similar situation.
This presentation will describe the experiences of a large, two and
one half year, outsourcing contract - the analysis of more than 16,000
sexual assault kits covering the years 1989 - 1998. These sexual assault
kits had been stored by the New York Police Department; in 2000 the
decision was made by New York City to contract out the analysis of
these sexual assault kits to three vendor laboratories. The New York
* Presenting Author
City Office of Chief Medical Examiner’s Department of Forensic
Biology, which did not have the responsibility to analyze sexual assault
cases prior to 1999, participated in the contracting process. As a result,
the Department of Forensic Biology took on many responsibilities of the
contract, including reviewing the STR data generated by the contract
labs for inclusion in CODIS.
There were many trials and tribulations encountered: setting up the
contracts; working out the initial bugs; preparing “dummy” sexual
assault kits for quality control purposes; developing methods of efficiently reviewing the data; confirming the performance of appropriate
positive and negative controls; checking the results obtained on the
“dummy” sexual assault kits; identifying and remediating problems in
the vendor labs; tracking the progress of shipments; identifying local,
state, and national database hits; notifying the necessary agencies; and
appearing in court proceedings.
The presentation will also describe some of the interesting outcomes of the project, including the identification of well over three
hundred sexual assault patterns of two cases or more and the freeing of
a man incorrectly convicted of a sexual assault in 1992.
Part II of this presentation is a poster presentation which will
present statistical information with regard to the analysis results, quality
of the data obtained, number of hits generated, etc.
DNA, Sexual Assault Kits, Outsourcing
B134 Degenerate Oligonucleotide-Primed PCR:
A Whole Genome Amplification
Approach to Forensic DNA
Kristin M. Meyer, BS, George Washington University, Washington, DC
20052; Kristen E. Lewis, BS, Virginia Commonwealth University, 1000
West Cary Street, Richmond, VA 23284; Moses S. Schanfield, PhD,
George Washington University, Washington, DC 20052; Tracey C.
Dawson, PhD*, Virginia Commonwealth University, 1000 West Cary
Street, Box 842012, Richmond, VA 23284
These research findings will provide a complete set of validated
protocols for the final optimized whole genome amplification experiments. All procedures will be developed for instrumentation and
equipment that currently exists in most forensic DNA laboratories.
These will impact the forensic DNA community by allowing for
increased success rate for cases which contain compromised biological
samples without a significant increase in costs or the need for specialized
training.
The goal of this research project is to provide the Forensic DNA
community with a tool – Whole Genome Amplification – that than can
readily increase the success rate of the analysis of degraded, aged or otherwise compromised biological evidence samples using existing conventional lab technologies and standard procedures for data analysis.
This presentation is aimed to familiarize the audience with one particular
method, DOP-PCR, for nuclear STR amplification and analysis.
Whole genome amplification techniques such as primer extension
preamplification (PEP), multiple displacement amplification (MDA),
and degenerate oligonucleotide-primed PCR (DOP-PCR) have been utilized in a variety of scientific areas, including embryonics, cancer
genetics, histopathology, and genetic disease diagnosis and linkage
studies. Recently, DOP-PCR has been successfully used as a method to
generate larger amounts of DNA that are necessary to perform SNP and
microsatellite genotyping. DOP-PCR involves the use of a 16-mer
degenerate primer which theoretically allows the statistical and representative amplification of the entire genome and has been shown to
accomplish this from as little starting material as that contained within a
single cell. This study focuses on using DOP-PCR to analyze forensic
samples where the yields of high quality DNA are too low for genotyping using conventional nuclear DNA methods. A sensitivity experiment was performed using genomic DNA to evaluate the technique’s
* Presenting Author
lower limits of amplification. Initially, input DNA amounts ranging from
1 nanogram to 15 picograms were tested using the recommended 100ul
amplification volume. In addition, mixed stain sperm fraction DNA
samples from known male and female contributors (representing “compromised case” samples) that had previously failed to generate a profile
were tested in order to approximate DOP-PCR’s ability to generate an
accurate genotype profile. Preliminary results indicate that DOP-PCR is
capable of increasing the amount of total DNA, particularly for lower
input DNA amounts. The total yield on average increased by several
hundred-fold at the lower dilutions using the DOP-PCR method.
However, the fragment sizes obtained after DOP-PCR indicate that
larger fragments are not amplified as successfully as smaller fragments.
The majority of DNA products resulting from DOP-PCR fell within a
500 bp to 1 kb range, which is slightly shorter than anticipated from published data reporting fragment sizes up to 3 kb. Additionally, the results
for the “compromised case” samples suggest that the DOP-PCR
products can successfully produce a correct profile with distinct and balanced peaks at STR loci. On average, Profiler Plus amplification of these
samples produced balanced peaks for 7 out of the 10 loci, with unbalanced or failed amplification resulting mostly in the loci with larger
expected fragment sizes. Further research will concentrate on the sensitivity aspect of this technique and the capability of obtaining larger
amplicons from limiting or highly degraded forensic samples for both
nuclear and mitochondrial genome applications.
DNA, STR, Whole Genome Amplification
B135 A Review of Fluorescent Artifacts
in Genetic Analysis Systems
Rhonda K. Roby, MPH*, Chu-An Chang, PhD, Natasha Coyle, PhD,
Dana Elliott, BA, Paul A. Foxall, PhD, Grace Lee, PhD, and Kathy
Wang, MS, Applied Biosystems, 850 Lincoln Centre Drive, M/S 404-3,
Foster City, CA 94404
After attending this presentation, attendees will have reviewed the
artifacts visualized with the use of the AmpFLSTR® PCR Amplification
Kits and ABI PRISM® genetic analysis instruments.
This presentation will support the caseworking forensic scientist in
further developing interpretation and troubleshooting skills.
Molecular biology techniques using DNA have revolutionized the
field of forensic sciences in testing of biological specimens. The
forensic community realized the advantages of DNA over traditional
serological techniques and first adopted restriction fragment length polymorphism (RFLP) and sequence-specific oligonucleotide (reverse dot
blot) assays. Many laboratories adopted silver-stained slab gels with
long tandem repeats following those techniques. Then, fluorescent dye
technology was introduced. Fluorescent multicolor dye technology
allows multiple loci (including loci that have alleles with overlapping
size ranges) to be analyzed in a single capillary injection or gel lane.
Alleles with overlapping loci are distinguished by labelling locus-specific PCR primers with different colored dyes. Laboratories can now
analyze hundreds of loci in a single day using five-dye fluorescent
labelling from Applied Biosystems (Foster City, CA). Additionally, the
forensic community has adopted the use of multicolor sequencing for
mitochondrial DNA analysis.
PCR-based technology, and especially current fluorescent short
tandem repeat (STR) analysis, has proven its advantages in the past
decade. PCR-based testing oftentimes produces results when the DNA
is highly degraded, unlike RFLP testing. PCR-based STR testing produces discrete results. PCR-based testing has much greater sensitivity
and the small size of the STR loci improves the chance of obtaining a
result, particularly for degraded DNA samples. Additionally, the small
size range of STR loci makes them ideal candidates for co-amplification
where multiple STR loci or other markers can be profiled in a single
amplification. Furthermore, advances in technology have resulted in an
89
increase in sensitivity, allowing the forensic community to obtain results
using far less input DNA than in previous years.
Forensic DNA testing for the identification of evidentiary material
involved in the resolution of legal disputes is a powerful technique.
With any scientific procedure, a quality assurance (QA) program that
addresses the techniques is critical. The forensic community has
excellent QA programs to assure accurate results and to meet the rigors
of the courts. Each method has its own advantages, limitations, and
quality assurance and quality control procedures. A good QA program
encompasses as many aspects of DNA testing as reasonably possible.
Once a sample is amplified for STR analysis using multiplex
AmpFLSTR® PCR Amplification Kits (Applied Biosystems) and run on
the ABI PRISM® genetic analysis instruments (Applied Biosystems),
many steps are performed and an array of reagents and consumable
items are used in the final analysis of that sample. Because of this, troubleshooting can be complex.
Inherent with all of these techniques and procedures are artifacts, or
anomalies. The presence of artifacts can be attributed to a number of
factors including phenomenon innate with the technique, cleaning procedures, and raw material. For example, with RFLP analysis, autoradiographs often show anomalies from static electricity, defective intensifying screens [Benzinger et al. “An illustrated guide to RFLP troubleshooting,” Journal of Forensic Sciences 1998; 43(3)665-679],
shadow bands, and fingerprints. The results obtained from samples
amplified for STR analysis using any one of the numerous fragment
analysis systems can exhibit their own anomalies. These anomalies can
be reproducible anomalies and non-reproducible anomalies. Examples
of these two (2) categories of anomalies will be explored.
Peaks other than the target alleles may be detected on the electropherogram displays. Examples of reproducible anomalies include stutter,
incomplete 3’ A nucleotide addition (-A), mixtures, and dye-labelled
artifacts. Artifacts can be intermittent and are not always reproducible.
In our experience, non-reproducible artifacts can be correlated to sources
other than the amplification chemistry or the moiety of the sample. For
example, spikes caused by salt accumulation or dried polymer released
from a dirty block and traveling through a capillary are examples of nonreproducible artifacts. Another example is an artifact resembling a
“stair-step” that may be associated with a need to change the water in the
autosampler tray on the ABI PRISM® 310 Genetic Analyzer.
Several artifacts associated with practices using the AmpFLSTR®
PCR Amplification Kits and run on the ABI PRISM® genetic analysis
instruments will be presented.
ABI PRISM and its Design, AmpFlSTR, and Applied Biosystems
are registered trademarks of Applera Corporation or its subsidiaries in
the U.S. and certain other countries. AB (Design) and Applera are trademarks of Applera Corporation or its subsidiaries in the U.S. and certain
other countries. All other trademarks are the sole property of their
respective owners.
Low copy number (LCN) analysis is an approach that involves the
examination of minute quantities (i.e., <100 pg) of DNA template. Even
though genetic profiles can readily be obtained from body fluid stains
when enough DNA is present for analysis, there are times when a genetic
profile cannot be obtained because of the presence of too little DNA for
analysis. When minute quantities of DNA are encountered in casework,
LCN typing can provide a means by which a genetic profile can be
obtained. In an attempt to provide casework laboratories with strategies
to increase their DNA typing success rate by being able to routinely
employ LCN methods, various LCN typing strategies were examined.
Initial experiments concentrated on the ability of increasing the
PCR cycle number, using the D1S80 PCR-VNTR system as a model, in
an attempt to type single or few cells. Cycle numbers ranged from the
standard 30 cycles up to 50 cycles. Results indicate that increasing the
cycle number to 35 cycles often resulted in the ability to type single or
few (<5) cells. Increasing the cycle number beyond 35 resulted in the
formation of non-specific amplification products that may obscure the
presence of the true alleles. Concomitant with the ability to type single
or few cells, a loss of heterozygosity (allelic drop-out) was observed.
Several whole genome amplification (WGA) methods were evaluated for LCN analysis in order to permit increased sensitivity of
analysis down to a singe cell equivalent. The WGA methods tested were
primer extension PCR (PEP), degenerate oligonucleotide primed PCR
(DOP), and multiple displacement amplification (MDA). PEP uses a
mixture of random 15 base oligonucleotide primers to prime and subsequently amplify the whole genome (or a large percentage of it) prior to
subsequent genetic analysis while DOP uses a single degenerate primer.
MDA is an isothermal process that employs random hexamer primers to
amplify the whole genome.
Cell suspensions were first subjected to WGA analysis using PEP,
DOP, and MDA. Aliquots of the products were then re-amplified using
primers specific for the D1S80 PCR-VNTR locus. Cycle numbers
ranged from the standard 30 cycles to 50 cycles. The results obtained
are promising. It appears that WGA products from PEP and DOP can be
used to increase the sensitivity of D1S80 analysis from a few cells to one
cell-equivalent. Results indicate that WGA products from MDA can be
used to type a few (<5) cells, but the sensitivity of D1S80 analysis is not
greatly increased using this WGA method. Allelic drop-out was
observed with each WGA method tested—PEP, DOP, and MDA—as
expected. Allelic drop-in and non-specific amplification products were
also observed. However, the extraneous alleles that were detected did
not interfere with genetic typing of the samples. The preceding WGA
analyses used the D1S80 PCR-VNTR system as a model; the WGA
assays are being applied to autosomal and Y-STR typing systems.
Detailed results will be presented including an evaluation of the efficacy
of these methods for casework specimens.
Low Copy Number (LCN), Increased Cycle Number (ICN),
Whole Genome Amplification (WGA)
Artifacts, Fluorescent, STR
B136 Strategies for Low Copy Number (LCN)
DNA Analysis
Virginia L. Raker, BS*, Erin K. Hanson, MS, and Jack Ballantyne,
PhD, National Center for Forensic Science, University of Central
Florida, Department of Chemistry, PO Box 162367, Orlando, FL
32816-2367
After attending this presentation, attendees will understand various
strategies that would enable samples containing low copy numbers of
starting DNA template to be DNA profiled
The forensic community may be able to use the LCN typing
strategies presented to obtain DNA profiles from samples containing low
copy numbers of starting DNA template.
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B137 SWGDAM Validation of a 19 Locus
Y-STR System for Forensic Casework
Darlene Daniels, MS* and Jack Ballantyne, PhD, University of
Central Florida, 12354 Research Parkway, Room 225, Orlando,
FL 32826; Ashley Hall, MS, National Center for Forensic Science,
PO Box 162367, Orlando, FL 32816-236
After attending this presentation, attendees will be presented with a
full SWGDAM developmental validation of two Y-STR multiplexes,
and will understand the unique requirements of such systems.
This presentation will impact the forensic community by demonstrating that few Y-STR multiplexes have undergone a full SWGDAM
validation. We intend to present the community with a complete set of
experiments and discuss the differences between the newly developed
systems and the current autosomal systems in use.
* Presenting Author
A SWGDAM developmental validation study was carried out on
two Y-STR multiplex systems (MPI and MPII) that collectively permit
the co-amplification of nineteen Y-STR markers including DYS393,
DYS392, DYS391, DYS389I, DYS389II, Y-GATA-A7.2, DYS438,
DYS385a and DYS385b (MPI); DYS425, DYS388, DYS390, DYS439,
DYS434, DYS437, Y-GATA-C.4, Y-GATA-A7.1, Y-GATA-H.4, and
DYS19 (MPII). Performance checks subsequent to PCR parameter optimization indicated that MPI and MPII were suitably reproducible,
precise and accurate for forensic use. The sensitivity of the systems was
such that a full 19-locus Y-STR profile was obtainable with 150-200 ng
of male DNA, and some loci were detectable even with as little as 20-30
pg of input DNA. Primate specificity was demonstrated by the lack of
cross reactivity with a variety of commonly encountered bacterial and
animal species, with the single exception of a monomorphic canine
product that was outside of the size range of human alleles from any of
the nineteen loci. Unsurprisingly cross reactivity was observed with a
number of male and female non-human primates. Environmentally
compromised samples produced full or partial Y-STR profiles. For
example, a semen stain exposed to the outdoor elements for six months
still gave a 13-locus Y-STR profile. Although a limited number of
female DNA artifacts were observed in mixed stains in which the male
DNA comprises 1/300 of the total, the full 19-locus male profile was
easily discernible. Even at a 1500-2000 fold dilution of male DNA with
female DNA partial Y-STR profiles were obtained. Furthermore the
potential utility of MPI and MPII for forensic casework is exemplified
by their ability to dissect out the male haplotype in a variety of case type
samples including, inter alia, post-coital vaginal swabs, admixed male
and female bloodstains, the non-sperm fraction from a differentially
extracted semen stain and to determine the number of male donors in
mixed semen stains.
SWGDAM Validation, Y-STR Multiplex, MPI/MPII
B138 Development and Characteristics
of a Novel Y-STR Multiplex PCR
Amplification System
Paul A. Foxall, PhD*, Sulekha Rao, PhD, Heidi Philips, and Robert
Stern, Applied Biosystems, 850 Lincoln Centre Drive, Foster City,
CA 94404
characteristics of a Y-STR multiplex amplification system suitable for
use in forensic analysis of samples containing mixtures of male: female
DNA.
This talk will acquaint the forensic community with a novel Y -STR
multiplexed kit that will provide reliable and robust Y haplotypes from
forensic samples. This will be able to be used in conjunction with their
existing instrumentation currently used for autosomal STR analysis.
Analysis of the Y chromosome is useful for tracing human evolution through male lineages and in a variety of paternity and forensic
applications. In a forensic setting, Y chromosome tests provide the
ability to separate and analyze the male DNA component from samples
containing mixtures of female and male DNA. If autosomal short
tandem repeat (STR) markers are used, preferential amplification of the
major component of the mixture can mask the genetic profile of the
minor contributor. A multiplex PCR amplification system containing Y
– STR loci can enhance the detection of low levels of male DNA in these
types of samples. Y - STR loci show moderate levels of polymorphism
when compared to autosomal STR’s currently used in forensic analysis
due to the haploid nature of the Y chromosome. The addition of recently
described Y - STR loci to the European minimal haplotype and Scientific
Working Group on DNA Analysis Methods (SWGDAM) recommended
loci increases the possibility of obtaining sufficiently discriminative
haplotypes for use in forensic investigations.
* Presenting Author
A multiplex PCR amplification system in development at Applied
Biosystems has been designed to include the complete European
minimal haplotype and SWGDAM loci of DYS19, DYS385, DYS389I,
DYS389II, DYS390, DYS391, DYS392, DYS393, DYS438, and
DYS439, along with a number of additional loci chosen for their power
of discrimination and allele size range. To ensure no overlap between
allele ranges, loci are labeled with 6 - FAM™, VIC®, NED™ and PET®
dyes. Thermal cycling conditions are optimized for the GeneAmp®
PCR Systems, with subsequent amplified products being run on ABI
PRISM® genetic analysis instruments in conjunction with G5 and G5v2
dye sets or modules (Applied Biosystems).
Male DNA samples and female DNA samples were quantified by
using a real - time PCR quantification system, the Quantifilerä Y Human
Male DNA Quantification Kit, or the Quantifiler™ Human DNA
Quantification Kit respectively (both Applied Biosystems, currently
under development at the time of writing this abstract). These reactions
were amplified and analyzed on an ABI PRISM® 7000 Sequence
Detection System (Applied Biosystems). From these data, the quantity
of amplifyable DNA in each sample was calculated and this value used
to prepare test samples. These test samples included serial dilutions
ranging from 2ng to 0.0625ng, along with male:male mixtures in ratios
of up to 10:1, and female:male mixtures in ratios of up to 200:1. Using
these samples, the amplification conditions were optimized for signal
strength, color balance and to minimize stutter and –A artifacts. Cross
reactivity studies showed no consistent peaks for male animals and
prokaryotes, although some reproducible peaks were seen with DNA
samples from higher primates. Haplotypes produced from the male
DNA samples were consistent across thermal cyclers and instruments
used throughout the development of the kit.
This Y - STR PCR amplification system and the Quantifiler™
Y Human Male DNA Quantification Kit, used in conjunction with
instrumentation from Applied Biosystems, are designed to produce
reliable and accurate Y - haplotypes and provide the forensic scientist
with a robust set of tools for Y - chromosome analysis.
For Research, Forensic or Paternity Use Only. Not for use in diagnostic
procedures. The PCR process and 5’ nuclease process are covered by
patents owned by Roche Molecular Systems, Inc. and F. Hoffmann-La
Roche, Ltd. ABI PRISM, Applied Biosystems, PET, VIC and GeneAmp
are registered trademarks and 6-FAM, NED and Quantifiler are trademarks of Applera Corporation or its subsidiaries in the US and/or certain
other countries.
®2003. Applied Biosystems. All rights reserved.
DNA Testing, Y-STR, PCR Amplification
B139 Examination of Non-Suspect Samples
Lacking Sperm Using a Y-STR 10-Plex
Cassie L. Johnson, MS*, Robert C. Giles, PhD, and Rick W. Staub,
PhD, Orchid Cellmark, 2600 Stemmons Freeway, Suite 133, Dallas,
TX 75207
The results of the study indicate that it is possible to obtain a male
STR profile even in the absence of spermatozoa by examining Y-chromosome STRs.
This study indicates that even in the absence of spermatozoa by
visual examination, it is possible to obtain a male STR profile by using
Y-STRs. Y-STR loci should become an important part of the CODIS
database in the future if they can be used in solving non-suspect sexual
assaults.
Y-chromosome short tandem repeats (Y-STRs) have gained interest
in the forensic community due to their ability to identify the male component of a sample. Y-STRs are particularly valuable in sexual assault
cases in which the amount of female DNA overwhelms the amount of
male DNA present, thereby making genotype interpretation more challenging. To detect male DNA from compromised sexual assault evi91
dence, ~45 non-suspect samples were analyzed with 10 Y-STRs. The
non-suspect samples were positive for the presence of human seminal
fluid, but were negative for the presence of spermatozoa by microscopic
examination. Complete or partial Y-STR 10-plex profiles were observed
in 27.6% and 52.9% of the samples, respectively. On samples yielding
partial profiles, results were obtained on an average of 5 loci per sample.
Approximately 19.5% of the samples did not yield any results. The
inability to obtain results may be due to either an insufficient amount of
amplifiable male DNA, PCR inhibition, or unfounded accusations of
sexual assault. The results of this study indicate that it is possible to
obtain a male STR profile even when there is no visible evidence of
spermatozoa. Furthermore, Y-STR loci should become an important
component of the CODIS database in the future if they are to be used in
solving non-suspect sexual assaults.
Y Chromosome, STRs, Forensics
B140 Detection of STR and SNP Markers
on the Y Chromosome Using the
Pyrosequencing Technology
Martina Nilsson, MSc*, Charlotte Johansson, MSc, Hanna
Andreasson, MSc, and Marie Allen, PhD, Uppsala University, Section
of Medical Genetics, Department of Genetics and Pathology, Rudbeck
Laboratory, Uppsala, SE 751 85, Sweden
After attending this presentation, the attendees will learn about the
principles of the Pyrosequencing technology and the possibility to detect
variations found on the Y-chromosome by using this flexible and rapid
method.This typing system is based on wellknown Y-SNPs and Y-STRs
that can be used in analysis of DNA found in degraded forensic or ancient
samples.
This typing system is based on a new technology which will improve
the analysis of short DNA fragments that are found in ancient samples and
in forensic casework materials. This technology detects variations found in
the genome and is very reliable and roubust.
Analysis of DNA sequence variation on the Y-chromosome is a useful
tool in forensic casework analysis, especially in cases of sexual assault
where mixtures of female and male DNA are found. Moreover, analysis of
Y-chromosome markers is valuable in studies of paternal relationships. The
routinely used Y-STR assay, which is based on determination of size after
fragment separation, is reliable and robust but will require analysis of large
fragments if the assay is multiplexed in order to analyze multiple markers
simultaneously. To be able to analyze ancient DNA and forensic casework
samples that have been exposed to severe environments resulting in degradation, amplification of short fragments is often necessary. We have
developed a system based on the pyrosequencing technology (SQA
analysis), which allow sequence analysis of short stretches of DNA to
detect SNP and STR variants found on the Y-chromosome.
Pyrosequencing is a non-electrophoretic, single-tube sequencingby-synthesis method, which is rapid in comparison to many other technologies used for SNP and STR analysis. The pyrosequencing assay is
flexible, easy to perform and will reveal the sequence of the repeats in
the STR rather than the length. Since the actual sequence is determined
in this assay additional information such as the nature of a mutational
event can be achieved. Moreover, the PCR fragments can be designed to
be very short, covering only a few bases outside the actual repeat unit,
to ensure a highly sensitive assay.
In this study, we have designed a typing system based on wellknown Y-STR and Y-SNP markers to be analyzed by pyrosequencing. A
major advantage with commonly used markers is that allele frequency
information from many different populations is available. A first set of
STR markers, DYS 392, DYS389-II, DYS438 and DYS 390 has been
evaluated. The PCR products for these markers yield fragments between
80 and 227 nucleotides. The repeat lengths for a number of male samples
were easily interpreted in the pyrosequencing analysis and all products
92
were male specific. A second set of markers, DYS19, DYS391,
DYS389-I and DYS393 is undergoing further development for pyrosequence analysis. The system also consists of 19 previously reported SNP
markers on the Y-chromosome, which are selected to be highly informative in Scandinavians. The primers for each fragment were designed
to be as short as possible, resulting in PCR products between 48 and 96
nucleotides. Specificity of the primers was first confirmed by evaluation
in singleplex PCR reactions, followed by analysis of the primers in multiplex PCR reactions to save valuable material.
In addition to the multiplexed PCR, time and reagents can be saved
by multiplex pyrosequencing analysis. A first combination of the STR
markers DYS392 and DYS438 have been analyzed simultaneously and
the repeat length of both markers can easily be inferred in the pyrogram.
The SNP markers have also been developed for performance of multiplex pyrosequencing. The SNP positions studied so far in these multiplexes were successfully interpreted and scored for the different polymorphisms.
In this study we have analyzed variations found on the Y-chromosome using the pyrosequencing technique. The use of short fragments
containing Y-SNP and Y-STR markers in this system will improve the
possibility to amplify and analyze degraded DNA in casework analysis
or ancient DNA studies. When fully developed analysis of teeth from
remains in a family grave, which were found 1915 in the church of
Varnhem, will be performed. These are the disputed remains of the
founder of Stockholm, Birger Jarl (1205-1266) and his son. Analysis of
Y-chromosome SNP and STR variation might support a paternal relationship between the two individuals in this case.
Y-STR, Y-SNP, Pyrosequencing Technology
B141 Real-Time Mitochondrial and
Nuclear DNA Quantification
of Forensic Evidence Materials
Martina Nilsson, MSc, Hanna Andreasson, MSc, and Marie Allen,
MD*, Uppsala University, Section of Medical Genetics, Department of
Genetics and Pathology, Rudbeck Laboratory, Uppsala, SE 751 85,
Swedzen
The attendee will learn about a new quantification technology
based on real-time 5´exonuclease detection, TaqMan. The system has
been used to determine the amount of DNA found in various evidence
materials. This technology can be used to choose the optimal target
(mtDNA or nDNA) and to avoid waste of valuable DNA material.
This TaqMan technology is a highly sensitive method that can
estimate the copy numbers of both mtDNA and nDNA simultaneously.
It is a quick and simple method that enables valuable DNA found in evidence materials not to be wasted.
Biological evidence material, found at a scene of a crime, often
contains limited amounts of DNA. Samples with scarce DNA amounts
are often analyzed by sequencing of mitochondrial DNA, due to its high
copy number per cell. Higher discrimination power is however obtained
by analyzing nuclear STR-markers, which is preferable when possible.
Since the choice of analysis method will be influenced by the amounts
of available DNA in a sample, a sensitive and accurate DNA quantification assay is essential in forensic DNA analysis. Moreover, DNA
quantification results can be used to estimate the optimal DNA amount
to be used in different experiments to ensure successful amplification
and avoid allelic dropout or preferential amplification.
We have developed a highly sensitive, rapid and reliable system for
quantification of nuclear and mitochondrial DNA copy numbers that
consumes a minimum of the valuable DNA sample. The system is based
on the real-time 5’exonuclease detection assay, using the ABI PRISM®
7700 instrument (TaqMan). Two specific probes, labeled with different
dyes, enables simultaneous quantification of the nuclear Retinoblastoma
1 gene and the mitochondrial tRNA Lys gene.
* Presenting Author
The quantification system has furthermore been used to determine
the DNA copy numbers available in a number of different evidence
materials frequently found at the scene of a crime. Analysis of cell debris
from different accessories, such as rings, watches and necklaces showed
large differences in DNA quantity. Shed hairs, roots from plucked hairs
and body hairs have also been quantified successfully using the quantification assay. Since shed hairs are common as evidence materials, differences in DNA quantity in the root part between hairs as well as within
single hairs at different lengths were studied. In addition, the high sensitivity and short amplicon length in the assay will allow DNA quantification of degraded and ancient DNA. To evaluate the assay performance
on ancient DNA it has been used for quantification of mitochondrial
DNA extracted from ancient bone remains found in a grave from the
1000-century in Sigtuna in mid Sweden.
This DNA quantification assay and the evaluation of DNA content
in different types of forensic materials have proven very useful in
forensic analysis. Moreover, it has been used to determine the minimal
amount of DNA required in several different DNA typing systems.
DNA Quantification, TaqMan Technology, Real-Time PCR
B142 Mitochondrial DNA Analysis
by Pyrosequencing
Susan Hastings, MS*, University of Central Florida, National Center
for Forensic Science, 12354 Research Parkway, Room 225, Orlando,
FL 32826; Jack Ballantyne, PhD, University of Central Florida, PO
Box 162367, Orlando, FL 32816-2367
The attendee will be introduced to mitochondrial DNA analysis by
pyrosequencing.
This presentation will introduce the community to mitochondrial
DNA analysis using a novel sequencing technology.
Mitochondrial DNA (mtDNA) analysis is often used in forensic
cases in which traditional nuclear DNA testing is, or is likely to be,
unsuccessful. Such cases typically involve compromised, or difficult-toanalyze specimens such as hair or bone. Currently, mtDNA analysis proceeds by standard sequencing of DNA isolated from such specimens.
Although current methodology is robust, its principal shortcoming is that
is is labor intensive and time consuming. This diminishes the efficacy
of mtDNA analysis in the investigative process and discourages its more
widespread use within the criminal justice system. We have evaluated a
new sequencing technology, pyrosequencing, for its potential applicability to forensic casework
Pyrosequencing uses the basic biochemistry of polymerase
mediated DNA chain elongation to determine the order of the bases.
When a deoxynucleotide triphosphate (dNTP) is added to the growing
DNA strand, a pyrophosphate (PPi) is released. A sulfurylase utilizes
this molecule to generate ATP, which in turn fuels a luciferase reaction.
Luciferin is converted to oxyluciferin, generating visible light proportional to the quantity of ATP. This light is detected by a CCD camera
and translated into a peak on the resulting program.
We have evaluated the performance of pyrosequencing with regard
to sensitivity, specificity, and compromised template DNA (including
mixtures). The advantages of this technology include vastly improved
timeliness of analysis (hours rather than days), a requirement of only
femtogram quantities of template DNA, and the potential for higher
mtDNA throughput by automation. This work has resulted in a set of
assays and associated standard operating procedures that should aid in
the transfer of the technology to operational casework and provide an
impetus for more widespread use of mtDNA testing for those cases
requiring it. These will be discussed in detail.
Mitochondrial DNA, DNA Sequencing, Pyrosequencing
* Presenting Author
B143 Real-Time mtDNA Specific Quantitation
Kerri A. Dugan, PhD, Helen Lawrence, MS, Mark Kavlick, BS, and
Elizabeth M. Olivastro, PhD*, Counterterrorism and Forensic Science
Research Unit, Laboratory Division, Federal Bureau of Investigation,
Quantico, VA 22135; Constance Fisher, PhD, DNA Analysis Unit II,
Laboratory Division, Federal Bureau of Investigation, Quantico,
VA 22135
After attending this presentation, attendees will learn by incorporation
of a mtDNA control region TaqMan assay into mtDNA sequence analysis
could be extremely beneficial to the forensic community.
A mtDNA control region TaqMan assay could reduce pre-amplification assay time and enhance casework results which is desirable because
forensic mtDNA testing is a lengthy, tedious and labor-intensive procedure.
Many laboratories that perform forensic mitochondrial DNA
(mtDNA) analysis use quantitation of nuclear DNA to estimate the
amount of mtDNA present in a DNA extract. Since mtDNA can be analyzed successfully from samples with little to no detectable nuclear
DNA, extracts are routinely processed even if the nuclear DNA quantitation is inconclusive. In addition, the slot blot hybridization technique
that is currently employed by many laboratories to quantitate extracted
DNA is time-consuming and relies on subjective interpretation of the
quantity of DNA. Alternatively, quantitative real-time PCR provides a
rapid, objective estimate of DNA quantity and is amenable to
automation. Importantly, quantitative real-time PCR provides the opportunity for custom quantitation. The TaqMan assay uses oligonucleotide
probes that hybridize to specific DNA sequences. The probes are
designed with a reporter fluorophore on the 5’ end and a quencher fluorophore on the 3’ end. During PCR, the probe hybridizes to the template
DNA downstream of the primer and the 5’ to 3’ exonuclease activity of
the DNA polymerase separates the reporter molecule from the quencher
molecule during template extension. Cleavage of the reporter molecule
results in a measurable increase in fluorescence. As each DNA strand is
extended an increase in fluorescence is observed. This presentation will
describe the development of a TaqMan based assay for quantitation of
mtDNA.
While others have described real-time PCR assays for quantitation
of mtDNA, these assays have targeted areas of the coding region of the
mtDNA genome. Previous work by our group has produced a real-time
PCR based assay for quantitation of mtDNA based on amplification of
region HVIA within the control region. As an extension of our previous
project, this work focuses on another, less polymorphic, mtDNA target
that lies in the control region of the mitochondrial genome. Since
forensic DNA analysis requires human specific DNA quantitation, it is
necessary to choose a target region that is conserved among humans but
does not amplify DNA from other species. Primers and probes were
designed to interrogate a region between HVI and HVII and are specific
to human or higher primate mtDNA.
Amplification reactions were optimized for amplification temperature, MgCl2 concentration, primer concentration and probe concentration. Once the conditions for optimal amplification were determined,
sensitivity and reproducibility studies were carried out under these conditions. The HL60 mtDNA control region was cloned into pCR2.1
(Invitrogen, Carlsbad, CA) to create a reagent for use as a mtDNA
standard to prepare standard curves. This allows the user to compare the
fluorescence generated by the sample to a standard curve and determine
the amount of mtDNA present in the extract. In some DNA extracts,
quantitation by methods such as slot blot hybridization reveals that DNA
is present but the DNA does not amplify under standard amplification
procedures. Two possible causes for this observation could be DNA
degradation or the presence of PCR inhibitors in the extract. The ability
to distinguish between degraded DNA and PCR inhibition has been
explored. In addition, the effect of various PCR inhibitors on the ability
to quantify mtDNA using real-time PCR has been studied. Finally, a validation study on evidentiary-type samples is underway.
93
Improvements that reduce assay time and enhance casework results
are desirable because forensic mtDNA testing is a lengthy, tedious and
labor-intensive procedure. Incorporation of a mtDNA control region
TaqMan assay into mtDNA sequence analysis could be extremely beneficial to the forensic community. This work has generated a reagent that
allows easy creation of standard curves for quantitation of mtDNA by
PCR as well as a TaqMan based quantitative PCR assay to quantify
mtDNA. This protocol has been optimized and the sensitivity and reproducibility of the method as well as the effect of PCR inhibitors on the
assay have been assessed. Finally, a study to validate this assay for use
in pre-amplification mtDNA quantitation has been undertaken.
Mitochondrial DNA, Pre-Amplification Quantitation,
Real-Time PCR
B144 Development of Microchip-Based
Sample Processing Systems for
Forensic DNA Analysis
James P. Landers, PhD* and Jerome P. Ferrance, PhD, University of
Virginia, Departments of Chemistry and Pathology, PO.Box 400319,
This talk will discuss the development of modular and integrated
microchip-based technologies that can be utilized in expediting sample
processing and analysis steps for forensic DNA analysis
The impact of this talk will be to explore the developments in
microfluidic processing and analyses which are applicable to forensic
DNA analysis.
Forensic DNA analysis is a multistep process that involves a
diverse array of molecular and analytical techniques executed in a
sequential manner with STR analysis as the final step. Individually,
these steps are time-consuming and labor-intensive, which has resulted
in a slow throughput rate for forensic analysis and contributed to the
backlog of samples awaiting forensic DNA testing. One approach for
reducing analysis time for the individual processes involved in forensic
DNA analysis is to exploit miniaturized technologies. In particular,
microfluidic devices have been utilized in biochemical processes and
analyses that are applicable to forensic work, with the added benefit that
they can be designed with monotasking functionality, or in an integrated
format for multiple process execution on a single device.
The last decade has seen an explosion of efforts to miniaturize
sample preparation steps, such as DNA extraction and PCR amplification, with the expressed goal being to expedite the molecular diagnostic evaluation of human samples. These efforts have demonstrated,
unequivocally, that the microminiaturization of analytical processes on
microchip platforms can lead to enhanced efficiency and a reduction in
time (versus conventional methods). In addition, optimized microchip
sample processing can accommodate small sample sizes and does so
with minimum reagent consumption. This has been demonstrated with
the microchip-based extraction of nucleic acids from complex samples
including whole blood, PCR amplification of specific genomic targets in
volumes (as low as a few hundred nanoliters) on microchips, microchip
electrophoretic separations for DNA analysis, and, to a lesser extent,
with chip-based sorting of blood cells in labyrinth-like silicon structures.
The power of analytical microchip technology for carrying out analytical processes rapidly and efficiently can be exploited in numerous
ways by the forensic community. Traditional forensic DNA analysis
methods, such as differential extraction, DNA purification and quantitation, STR amplifications, and capillary electrophoretic separations,
can be replaced with equivalent microchip methods that provide a equivalent, if not higher, level of confidence. One embodiment of a forensic
microdevice design involves a ‘modular’ approach, which aims to
develop stand-alone, dedicated instrumentation that accepts single-task,
single use microchips designed for automated analysis. These are being
94
developed for cell sorting, DNA extraction, and PCR amplification
processes. These microdevices are designed in a ‘task-conscious’
manner so that easy introduction into the conventional sample processing work-flow is possible. An alternative embodiment, the ‘integrated’ approach, is being pursued to create multi-task microchips
capable of carrying multiple processes in a sequential manner. This
requires the seamless interfacing of at least two different chip
microstructures, both fluidically and electrically, so that a single sample
can be processed through multiple steps automatically. Challenges here
are multidisciplinary in nature, requiring efforts in electrical and
mechanical engineering, surface science, polymer chemistry, molecular
biology and analytical chemistry to be brought to bear on the problem.
Ultimately, one can envision a forensic micro-total analysis system
(-TAS) that could accept a sample, extract and sort cells if required,
purify and quantify the DNA, amplify the target sequences of interest
and then electrophoretically separate the fragments with the single base
pair resolution required to generate an STR profile of forensic utility.
For rape kit evidence in particular, this type of microdevice processing
will provide male and female DNA analysis from a single swab in a reasonable amount of time, allowing faster evidence processing for current
cases, and contributing to reducing the significant backlog of cases
which currently exist.
Microchip Technology, Integrated Microdevices,
Sample Processing
B145 Automated PCR Setup for
Casework Samples as Part
of a Total Automated System
Arni S. Masibay, PhD, MSFS*, Paraj Mandrekar, MS, Laura
Flanagan, BS, Ryan Olson, BS, Michelle Mandrekar, MS, Robert
McLaren, PhD, and Allan Tereba, PhD*, Promega Corporation, 2800
Woods Hollow Road, Madison, WI 53711
After attending this presentation, the participant will have an understanding about a new automated PCR setup system designed for
casework samples that is flexible, customer friendly and integrated with
automated DNA purification and quantitation modules.
This presentation will impact the forensic community by demonstrating streamlined casework processing to alleviate the backlog cases
inundating the community.
Automated workstations have proven their utility for processing
sample backlogs for offender databases. These systems can be optimized
for a single sample type and do not have to be efficient. The varied
sample quality and amount encountered in casework presents additional
challenges in automated sample processing. Additionally, contamination
concerns restrict the flexibility of robotic systems and require extensive
validation. This presentation will discuss advances in developing an
automated PCR setup system that works over a 50 fold range of initial
DNA concentration. In addition, improvements for automated DNA
extraction and human-specific quantitation using the same robotic
instrument as PCR setup, the Beckman Coulter Biomek® 2000
Workstation, will be described. Developed initially as independent
modules, these three steps can be integrated with minimal hands-on
time. This modular approach provides quicker access to automation and
minimizes cost.
Automated PCR setup has been in use for many years. However,
due to the wide range of DNA concentrations associated with casework
samples, few laboratories have successfully automated this tedious and
time-consuming process. We modified the Beckman Coulter
Normalization Wizard developed for the Biomek® 2000 Workstation so
the program can dilute the DNA to a customer defined concentration
starting with DNA concentrations between 0 and 5ng/µl. The initial
DNA concentration values are imported from a modified AluQuant®
* Presenting Author
Calculator. The user then selects the final concentration and volume
parameters. The system will flag samples that are too dilute or concentrated and allow the user to exclude individual wells. This process conserves on expensive amplification reagents. After the DNA has been
diluted to the desired concentration a set amount of this DNA and PCR
master mix are added to a PCR plate and mixed. The user then caps the
plate and places it in a thermal cycler. Master mix can also be added
manually if desired.
We have integrated this PCR setup module with DNA purification
and quantitation on the same robotic platform to maximize its usefulness
and are continuing to gain experience on new sample types. To provide
the maximum recovery and flexibility, samples are currently preprocessed manually to remove biological material from solid supports.
In most cases, this involves incubation of the support in DNA IQ™ Lysis
Buffer, followed by centrifugation through a spin basket. Samples containing very small amounts of DNA are incubated in a Proteinase K
solution while samples containing sperm and epithelial cells are treated
using the standard differential extraction procedure. Once the samples
have been extracted from the solid support they are transferred to a
Biomek® 2000 Workstation for hands off purification using the DNA
IQ™ System. Recent modifications to this automation program reduce
processing time and adjust for environmental factors, such as low
humidity.
Human-specific quantitation is required for casework samples to
ensure the amplification of an appropriate amount of DNA. The
AluQuant® Human DNA-Specific Quantitation System was developed
to allow an automated approach to this step. Using solution-based
hybridization of a highly repeated human specific sequence, the method
is sensitive, provides numerical results, and does not rely on amplification of the sample DNA. Recent improvements to the automated
process allow the use of 4µl of sample DNA and improve sensitivity. In
addition, the DNA concentrations calculated in the AluQuant®
Calculator are automatically formatted for easy importation into the
Normalization Wizard program.
While not yet a “black box” sample analysis system, the current
setup provides a flexible system that automates several time consuming
processing steps on one robotic platform. Hands-on time between the
different programs is minimized and primarily involves replenishing the
deck with labware and reagents.
Automation, PCR, Quantification
B146 Miniplex Primer Sets: Sensitivity,
Peak Balance, Inhibitor and
Concordance Studies
Denise Chung*, Jiri Drabek, PhD, Kerry L. Opel, MA, and Bruce R.
McCord, PhD, Ohio University Department of Chemistry, Clippinger
Laboratories, Athens, OH
After attending this presentation, attendees will have been presented research on the current development of the Miniplex primer sets
for the analysis of degraded DNA.
This presentation will discuss the utility of the Miniplex primer sets
in the analysis of degraded DNA. In degraded DNA, only low concentrations of DNA template are generally available and PCR inhibitors
may be present. The effect of DNA template concentration on signal
intensity and peak balance ratio will be presented. This presentation will
also discuss the effect of PCR inhibitors on the amplification efficiency
of the Miniplex primer sets. Lastly, a full concordance study of 541
DNA samples as part of the method validation process will be presented.
In DNA recovered from the crime scene or site of disaster, the possibility of finding an intact target DNA sequence is greatly reduced due
to extensive fragmentation of the DNA template. Most commercial kits
have amplicon products ranging from 100-500 base pairs. Thus, in multiplex typing kits with a wide range of amplicon sizes, the larger PCR
* Presenting Author
amplicons often have lower signal intensity and fall below the detection
threshold. Re-designed Miniplex primers are positioned as close as possible to the ends of the stretch of repeats to reduce the amplified product
size. However, in situations where DNA is degraded, the DNA template
is not only highly fragmented but the concentration of the DNA template
recovered is also low. To test the hypothesis that shorter PCR amplicons
can improve amplification efficiency, we examined the effect of DNA
template concentration on signal intensity and peak balance ratio.
DNA concentrations ranging from 31 pg to 500 pg were amplified
with Miniplex 2, Miniplex 4 and Big Mini in 25 µl reaction volume. The
Miniplex primer sets were able to successfully amplify DNA targets at
these concentrations although these are below the range recommended
for commercial sets. For example, amplification at 100 pg/ 25 µl gave
signal intensities of 2000 RFU and 800 RFU for Miniplex 2 and
Miniplex 4, respectively. For the Big Miniplex, template concentrations
greater than 250 pg/ 25 µl were needed to avoid allele dropout. Good
peak balance ratio (>0.6) was achieved at these concentrations. Primer
concentrations for the Big Miniplex were further tested to improve the
sensitivity and peak balance ratio of this multiplex set.
Another challenge associated with forensic samples is the presence
of PCR inhibitors. The presence of these compounds can interfere with
the amplification process resulting in preferential amplification of one
locus, allele drop out, or no amplification at all. The effect on amplification efficiency of the Miniplex primer sets due to PCR inhibitors such
as hematin, indigo dye, humic acid, melanin, calcium, and collagen was
also investigated and will be presented.
Although all known polymorphisms have been taken into account
in the design of the Miniplex primers, primer binding related problems
may occur. Only comparison studies can verify the presence of previously undetected polymorphisms. A concordance study of 541 DNA
samples was performed to check for the existence of mutations that
could lead to allele dropout or low sensitivity of one allele in standard
STR typing kits. At the same time, potential point mutations in the
Miniplex primer binding region or insertion/deletion between commercial primers and Miniplex primers were also investigated. These
samples were obtained from the National Institute of Standards and
Technology (NIST) and were previously typed with the commercial kit
AmpFlSTR™ Identifiler.
Overall, these Miniplex primer sets can provide an alternative to
standard STR typing kits when allele drop out and low sensitivity of
large amplicons becomes a problem due DNA degradation, PCR inhibition or primer binding site mutations.
Degraded DNA, Miniplex, STR
B147 Development of New miniSTR
Loci for Improved Analysis of
Degraded DNA Samples
John M. Butler, PhD*, National Institute of Standards and Technology,
100 Bureau Drive Mail Stop 8311, Building 227, Room A243,
Gaithersburg, MD 20899-8311; Peter M. Vallone, PhD, National
Institute of Standards and Technology, 100 Bureau Drive, Mail Stop
8311, Gaithersburg, MD 20899-8311; Michael D. Coble, PhD*,
National Institute of Standards and Technology, 100 Bureau Drive,
Mail Stop 8311, Gaithersburg, MD 20899-8311
An approach to developing new DNA typing markers for improved
analysis of degraded DNA samples will be described.
The community will learn about new DNA tests that our group is
developing to improve analysis of degraded DNA samples. This will be
of interest to DNA analysts doing casework or thinking about handling
samples from mass disasters.
A number of studies have demonstrated that successful analysis of
degraded DNA specimens from mass disasters or forensic evidence
improves with smaller sized polymerase chain reaction (PCR) products
95
(1). Forensic DNA analysts often perform short tandem repeat (STR)
typing on highly degraded biological material and then turn to mitochondrial DNA testing, which is less variable but more likely to obtain
a result due to higher copy numbers in cells, if many or all of the STRs
fail. The commercially available kits for multiplex amplification of the
13 CODIS (FBI’s COmbined DNA Index System) STR loci usually
exhibit allele or locus-dropout for larger sized loci with degraded DNA
or samples containing PCR inhibitors.
By moving PCR primers closer to the STR repeat region, we have
demonstrated that it is possible to obtain fully concordant results to the
commercial kits while improving successful analysis of degraded DNA
with smaller PCR products or miniSTRs (1). However, many of the
CODIS core loci have large allele ranges (e.g., D21S11 and FGA) that
make it impossible to create small PCR products. Thus, we are going
beyond the CODIS core loci and examining a battery of new potential
STR loci that can be made less than 100 bp in size and would therefore
be helpful in testing highly degraded DNA samples. These new STR loci
are being put together into novel DNA testing assays and evaluated
across more than 600 samples representing the three largest populations
in the U.S.: Caucasian, African American, and Hispanic.
Methods and Materials: Following selection of potential new STR
loci, PCR primers were designed using a standardized approach that has
been described previously (2). For the miniSTR loci, a single locus is
placed into each dye color in order to keep the size ranges less than 100
bp where possible (1). Candidate loci were selected from STRs located
on chromosomes that did not possess any CODIS STRs (i.e., chromosomes 1, 6, 9, 10, 14, 15, 17, 19, 20, 22, and X) so that results could be
treated as unlinked and the product rule employed in any statistical
analysis between the CODIS loci and new ones.
Summary of Results: New PCR primers were designed and tested
for a dozen new STR loci. All loci show a moderate degree of polymorphism in the U.S. population samples tested and compared well to results
from the CODIS loci in the same sample set.
Conclusions: The selection of STR loci that have a narrow allele
size (e.g., less than 50 bp) and can be made smaller than 100 bp works
well with degraded DNA samples. In addition, these new STR loci that
are under development will be helpful in analysis of casework involving
close relatives including complicated forensic paternity cases (e.g.,
incest) where the 13 CODIS loci are not enough.
(1) Butler, J.M., Shen, Y., McCord, B.R. (2003) The development of
reduced size STR amplicons as tools for analysis of degraded DNA. J.
Forensic Sci., in press.
(2) Schoske, R., Vallone, P.M., Ruitberg, C.M., Butler, J.M. (2003)
Multiplex PCR design strategy used for the simultaneous amplification
of 10 Y chromosome short tandem repeat (STR) loci. Anal. Bioanal.
Chem., 375: 333-343.
Short Tandem Repeat DNA Typing, Degraded DNA, Reduced Size
PCR Products
B148 mRNA Profiling: Body Fluid Identification
Using Multiplex RT-PCR
Jane S. Juusola, BS*, University of Central Florida, Department of
Chemistry, PO Box 162366, Orlando, FL 32826-2366; Kevin W.P.
Miller, PhD, FBI Academy, Building 12, CTFSRU, Quantico, VA
22135; Jack Ballantyne, PhD, National Center for Forensic Science,
University of Central Florida, PO BOX 162367, Orlando, FL
32816-2367
After attending this presentation, attendees will have been presented with a novel means of identifying body fluids of forensic interest.
The forensic community will be shown how the highlighted system
could supplant the battery of serological and biochemical tests currently
employed in the forensic serology laboratory.
96
Since it can be important to identify the nature of the body fluids
present in a stain recovered at a crime scene, the development of a body
fluid identification system that is compatible with current DNA typing
procedures is desirable. Conventional methods of body fluid identification use labor-intensive, technologically diverse techniques that are
performed in a series, not parallel, manner and are costly in terms of time
and sample. Moreover, for some frequently encountered body fluids no
confirmatory technique exists. There is no definitive test, for example,
for the presence of saliva or vaginal secretions. In seeking to develop
novel multiplex (i.e., parallel) analysis procedures for body fluid identification that are compatible with current DNA analysis procedures, we
have considered assays based upon protein and messenger RNA
(mRNA) since both are expressed in a tissue specific manner. However,
multiplex analysis of complex, partially degraded protein mixtures such
as those present in body fluid stains awaits further developments in proteomics. Messenger RNA is considered a better option because the technologies for massively parallel analysis continue to be developed due to
the rapidly evolving field of functional genomics.
Terminally differentiated cells, whether they comprise of blood
monocytes or lymphocytes, ejaculated spermatozoa, or epithelial cells
lining the oral cavity become such during a developmentally regulated
program in which certain genes are turned off whereas others are turned
on. Thus, a pattern of gene expression is produced that is unique to each
cell type, which is evinced by the presence and relative abundance of
specific mRNA species. If the type and abundance of mRNAs could be
determined in a stain or tissue sample recovered at the crime scene it
would be possible to definitively identify the tissue or body fluid in
question. Advantages of an mRNA-based approach, compared to conventional biochemical analysis, include greater specificity, simultaneous
and semi-automatic analysis though a common assay format, improved
timeliness, decreased sample consumption and compatibility with DNA
extraction methodologies.
Previously we have reported that it is possible to isolate total RNA
of sufficient quality and quantity from biological stains to enable subsequent detection of particular mRNA species using the RT-PCR technique
and that we have identified candidate sets of saliva-, and semen-specific
genes. Since that time, we have also identified and tested candidate sets
of blood- and vaginal secretions-specific genes using a combination of
literature and public database searches.
In the extraction method that we employ, total RNA is isolated from
biological stains by extraction with guanidine isothiocyanatephenol:chloroform and precipitated with isopropanol. The extracted total
RNA is treated with DNase I, and then reverse-transcribed using random
decamers as the first strand primer. Finally, the cDNA is amplified using
gene-specific primers. The RT-PCR amplimer sizes are carefully chosen
to span the range of 100bp–350bp to allow facile separation on agarose
gels followed by visualization with a nucleic acid stain or by other platforms, such as capillary electrophoresis.
In the present work, we report the development of a multiplex
RT-PCR assay for the definitive identification of all of the body fluids
commonly encountered in forensic casework analysis, namely blood,
saliva, semen, and vaginal secretions. The tetraplex is composed of four
body fluid specific genes and has been optimized for the detection of
blood, saliva, semen, and vaginal secretions as single or mixed stains.
The methodology is based upon gene expression profiling analysis in
which the tissue specific genes are identified by detecting the presence
of appropriate mRNA species.
An mRNA based approach, such as the multiplex RT-PCR method
described above, could allow the facile identification of the tissue components present in a body fluid stain and conceivably could supplant the
battery of serological and biochemical tests currently employed in the
forensic serology laboratory.
mRNA Profiling, Multiplex RT-PCR, Body Fluid Identification
* Presenting Author
B149 Stability and Recovery of
mRNA in Biological Stains
Mindy E. Setzer, BS*, Jane S. Juusola, BS, and Jack Ballantyne, PhD,
University of Central Florida, PO Box 2367, Orlando, FL 32816-2367
The attend will learn about the persistence of mRNA in biological
stains.
This presentation will impact the forensic community by demonstrating mRNA is useful in determining the type of biological stain
present and persists under various environmental factors.
In theory, RNA expression patterns, including the presence and relative abundance of particular RNA species, provide cell and tissue specific information that could be of use to forensic scientists. An mRNA
based approach could allow the facile identification of the tissue components present in a body fluid stain and conceivably could supplant the
battery of serological and biochemical tests currently employed in the
forensic serology laboratory. Some of the potential advantages include
greater test specificity, and the ability to perform simultaneous analysis
using a common assay format for the presence of all body fluids of
forensic interest.
Previously we have reported that it is possible to isolate total RNA
of sufficient quality and quantity from biological stains to enable subsequent detection of particular mRNA species using the RT-PCR technique. In the extraction method that we employ, total RNA is isolated
from biological stains by extraction with guanidine isothiocyanatephenol:chloroform and precipitated with isopropanol. The extracted total
RNA is then treated with DNase I, at which time the extract can be quantitated, when desired, using a sensitive fluorescence assay based upon
the binding of the unsymmetrical cyanine dye RiboGreen. The RNA is
reverse-transcribed using random decamers as the first strand primer and
then the cDNA is amplified using gene-specific primers. We have
already identified numerous candidate body fluid specific genes for
blood, saliva, semen, and vaginal secretions using a combination of literature and public database searches.
To address concerns over stability of RNA in forensic samples, we
have conducted an in-depth study on the persistence of RNA in biological stains. Stains were prepared from blood, saliva, semen, and
vaginal secretions and exposed to a range of environmental conditions
so that affects of different light sources (UV, fluorescent, natural), temperature (room temperature, refrigeration, freezer), and environment
(outside covered, outside exposed) could be assessed. Initially, quantitation experiments were performed using biological samples of various
sizes (whole 50 ul stain, ¾ stain, ½ stain, ¼ stain, and 1/ stain) in order
8
to determine how much RNA can be recovered from the different sized
stains. We also performed RT-PCR with these stains using housekeeping
and tissue-specific gene candidates to determine sensitivity of the different primer sets. The results from these experiments were then applied
to the analysis of the stability samples which were collected at specific
time intervals (1 day, 3 days, 7 days, 1 month, 3 months, 6 months,
1 year, >1 year). The extent of RNA degradation within each type of
body fluid stain was determined using quantitation of total RNA and
reverse-transcription polymerase chain reaction with at least one highabundance and one low-abundance gene. DNA was also extracted from
the stability samples and quantitated to determine how the stability of
RNA compares to the stability of DNA in biological stains. The results
of these studies will be presented in detail.
RNA, Body Fluid Stain Identification, RNA Stability
* Presenting Author
B150 Forensic DNA Identification of
Feline Hairs: Casework and a
Mitochondrial Database
Joy L. Halverson, DVM, MPVM*, Questgen Forensics, 1902 East 8th
Street, Davis, CA 95616; Leslie Lyons, PhD, Department of Population
Health and Reproduction, School of Veterinary Medicine, University of
California, Davis, CA 94022
After attending this presentation, attendees will have a greater
awareness in the forensic community of the tools and utility of
individual identification of pets as part of criminal investigation. DNA
identification of animal hairs via mitochondrial haplotyping can provide
a useful link between a victim and a suspect.
This presentation will demonstrate that animal hairs are a common
finding in a carefully scrutinized environment such as a crime scene.
The application of forensic DNA analysis techniques to hairs and other
animal-derived samples opens previously unrecognized avenues of
criminal investigation.
DNA typing of samples from pets, including cats, has contributed
to homicide investigations and convictions in the United States and
Canada. Indeed the first example of forensic DNA identification of
animal hairs was a homicide investigation in Nova Scotia in which cat
hair was found in the pocket of a jacket discarded near the murder scene.
STR profiles of the hair matched that of Snowball, a cat belonging to the
suspect’s parents. While animal hairs are the most frequent animalderived sample recovered by crime scene investigators, such hairs are
usually shed telogen hairs.Despite the availability of feline STR
markers, there is often insufficient nuclear DNA to perform STR typing.
As in humans, mitochondrial DNA can be extracted from the hair shaft
and characterized by DNA sequencing or other sequence detection
methods. Assigning the significance of a DNA match between an
evidence sample and reference animal requires knowledge of the
frequency of the mitochondrial type in the domestic cat population. This
paper presents the database derived from DNA sequencing of the
mitochondrial control region from 155 purebred cats and 105 mixed
breed cats.
Domestic cats have elements of complexity in the mitochondrial
control not seen in dogs or humans. Between a relatively short Hv1
region and the Hv2 region is an AT-rich region with 3-5 tandem, 80 base
pair repeats. While this repeat region can be avoided by the amplification and sequencing of just the 3’ half of the control region, the 5’ end
is rich in polymorphism. Roughly 50% of cats actually show length
polymorphism when the entire control region is amplified. Two to three
fragments 80 base pairs apart can be electrophoretically separated
demonstrating that the variability is derived from the tandem repeat
region. A systematic analysis of this variability is required to ascertain
its utility in reliable individual identification.
In order to generate a feline mitochondrial database, primers were
designed to amplify the entire feline mitochondrial control region
(approximately 1100-1300 base pairs) analogous to 16000 bp to 400 bp
of the Andersen human mitochondrial genome. A total of 260 cats,
including 155 purebred cats representing 14 foundation cat breeds, as
well as 105 mixed breed cats were analyzed. The samples from the
purebred cats had been held in storage for some years and many were too
degraded to amplify the entire region. Sets of internal primers were
designed to amplify smaller, overlapping regions for sequencing. These
primers are also being investigated for nested PCR amplification, a procedure often required for processing of evidence hairs. Nested amplification through the tandem repeat region presents additional challenges
that are being investigated. A separate study has been undertaken to
ascertain whether heteroplasmy was detectable in feline hair samples
and, if so, to what degree. Results from the study are important for the
continued validation of feline mitochondrial haplotyping.
97
Feline mitochondrial typing was useful in a recent homicide investigation. In Iowa in 2000, Tracy Ann Carson disappeared; her body was
found 7 months later. The body had been wrapped in a large piece of
fabric, partially burned and buried; spring flooding unearthed it.
Investigators found a variety of animal hairs on the fabric. Feline hairs
taken from the fabric had mitochondrial haplotypes consistent with the
three cats owned by the suspect. Based on a previous database based
only on the 3’ end of the feline control region, the haplotype frequency
of the two siblings cats was 31% (the most common type) and the haplotype frequency of the third cat was 1.25%. Just before trial the suspect,
Ben O’Donnell, pleaded guilty to second degree murder.
DNA typing of animal-derived samples opens new possibilities for
linking suspects to crime scenes or victims. The close relationship between
people and their pets is a potentially valuable source of evidence to the
observant investigator. The value of such evidence will be determined by
the scientific validation of both its power and its limitations.
Animal Hairs, DNA Identification, Mitochondrial DNA
B151 DNA Profiling for Forensic
Soil Comparisons
Lorraine E. Heath, BSc* and Venetia A. Saunders, PhD, Liverpool
John Moores University, School of Biomolecular Sciences, Byrom
Street, Liverpool, L3 3AF, United Kingdom
After attending this presentation, attendees will have been
presented with the preliminary finding of a novel approach to differentiating soil samples for forensic applications.
This presentation will provide the forensicmcommunity a basis for
further research into the use of DNA profiling technology to identify
and/or compare forensic soil samples.
This paper will present the results of a preliminary investigation
that aimed to develop a reliable method of microbial DNA profiling to
differentiate between surface soil samples. Samples collected from given
locations with different ecological characteristics, such as uncultivated
fields, forests, and sand dunes were compared, and the variation within
and between ecologically different sites was determined. The implications of this research for forensic examination of soil evidence will be
discussed.
Forensic soil comparisons can be used to associate a person found
with soil on their clothes or possessions with a crime scene. Criminals
often commit crimes, or deposit evidence, in isolated areas such as fields
or forests. In most forensic cases where soil comparisons would be considered, a crime would have been committed in a known location (e.g.,
a field), and a suspect apprehended, who was found to have soil on their
clothes, shoes, or possessions (such as a shovel or vehicle tyres). It
would then be useful to compare these soil samples with control samples
collected from the scene to determine if the suspect’s samples could have
originated from that location.
Most current forensic soil comparisons are based on geological
properties. However, since the majority of forensic laboratories cannot
afford an expert geologist, such analyses are rarely performed for routine
casework, but are reserved for particularly high-profile, serious crimes.
It has been suggested that DNA profiling of the microbial community in
soil may be a potential method for soil comparisons. Currently,
microbial DNA profiling techniques such as Amplified Ribosomal DNA
Restriction Analysis (ARDRA), Denaturing Gradient Gel
Electrophoresis (DGGE), Thermal Gradient Gel Electrophoresis
(TGGE), Single Strand Conformation Polymorphism (SSCP), and
Terminal Restriction Fragment Length Polymorphism (TRFLP) are
being used to study the diversity of soil microorganisms. However, only
ARDRA and TRFLP can be performed on the equipment normally
available in the forensic laboratory. An ARDRA profile of a community
98
with many different species tends to become too complex for easy interpretation, as one species can contribute 4-6 restriction fragments.
However, TRFLP involves labelling the 5’ end of the PCR primers with
a fluorescent dye, so that only the terminal restriction fragments are
detected following gel electrophoresis. This means that TRFLP yields
less complex profiles where every visible band (fragment) represents a
single ‘ribotype.’ Additionally, TRFLP analysis relies on the detection
of fluorescently labelled primers, which is how human DNA profiling is
performed. Therefore, all necessary equipment and expertise should be
available in forensic laboratories, making this technique potentially
more applicable. Moreover, the technical nature of TRFLP would allow
high throughput of samples, as required for forensic applications.
As a pre-requisite for TRFLP analysis, various DNA extraction
methods suitable for a range of soil types were evaluated. An appropriate
procedure was developed for rapid and reliable extraction of DNA representative of the total microbial community. PCR was performed using
fluorescently labelled universal primers targeting the 16S ribosomal
RNA genes of the bacterial soil community. TRFLP analysis was then
performed using the ABI Prism 373 gel electrophoresis system and
GeneScan software. The resulting profiles were standardized and binary
code was assigned to the peak profiles. Similarities between these binary
codes were determined using the Jaccard coefficient to create similarity
matrixes and dendograms were used to display the results of agglomerative hierarchical cluster analysis.
Preliminary results suggest that samples from within a specific ecological site (e.g., a field) show a higher similarity to each other than to
those from other ecological sites (e.g., a forest). These results may allow
the determination of characteristic profiles that will facilitate identification of ecologically different sites so that a given sample collected
from a suspect could be identified as originating from, for example, a
field, rather than a forest. Further research will explore small-scale
spatial variation at various sites and the potential of these techniques to
link a suspect’s sample more precisely to its origin, and thus provide
stronger circumstantial evidence.
Soil, Microbial DNA Profiling, TRFLP
B152 Scientific Crime Scene Investigation
H. Dale Nute, PhD*, Florida State University, 4750 Collegiate Drive,
Panama City, FL 32405
After attending this presentation, attendees will have learned that
crime scene investigation both requires a scientific examination and can
meet the criteria mandated by the scientific method.
The most critical problem in forensic science today is the examination of crime scenes. It is the initial forensic science examination and
its results are the predicate for all the highly technical and sophisticated
examinations that follow. Any unreliability in the crime scene examination can seldom be rehabilitated by subsequent laboratory examinations. Proper investigation requires a scientific examination. This presentation lays the foundation for a scientific approach to crime scenes.
The foray initiated by the Daubert decision seems not to have
impacted crime scene examinations as it has the rest of forensic science.
In large part, this seems to be due to the prevailing opinion that crime
scene examinations are a technical activity, not a scientific one. There
are some, however, who advocate that crime scene examinations not
only are included in the overall discipline of forensic science but are the
fundamental examination. There is little question but that it is the initial
examination and that the reliability of all subsequent examinations
depends on the quality of the evidence collected at the scene. But
merely being the precursor for a scientific examination does not make it
scientific. This presentation will advocate that the nature of a crime
scene examination requires that it be performed scientifically and delineates the criteria that it must meet.
* Presenting Author
There are those who accord scientific stature only to disciplines
based on technology. Science, however, is as much a thought process as
a testing process. A scientific examination must have be based on a valid
theory, conducted according to a reproducible, empirical protocol, and
evaluated according to objective decision criteria. These criteria apply to
crime scene examinations.
There are those who accord scientific stature only to endeavors that
have a corresponding academic discipline. The investigation of
evidential materials associated with crimes and accidents requires a
wide-ranging set of background knowledge and skills incorporating
aspects of most of the scientifically based forensic science disciplines. In
particular, background knowledge touching on engineering (vehicular
accidents and materials science), medicine (cause and manner of trauma
and death), and criminology (criminal motivation and conduct of crime)
is required. Obviously, one must have a sound grounding in the basic
sciences of chemistry, physics and biology, as well as mathematics,
logic, and statistics, to conduct competent investigations of evidential
materials.
There are those who would deny scientific status to crime scene
investigations because they are performed by non-scientists, (i.e., investigators). This is similar to the argument used against some of the other
forensic sciences and is fallacious. The two major consideration of a
crime scene investigation are reliability and relevance. The characteristics of a scientist are required for reliability – conducting valid, reproducible and objective examinations to obtain and interpret data. The
characteristics of an investigator are required for relevance – applying
inferences from the data obtained in order to solve crimes.
Unfortunately, because our society is split into a dichotomy of scientists
and non-scientists, mirrored by a dichotomy of forensic scientists and
investigators, this combination investigator/scientist currently is a rarity.
It does not have to remain so.
The distinguishing characteristic between a technician and a scientist is that the scientist interprets evidence while a technician only collects evidence. To begin the educational process to upgrade crime scene
technicians into crime scene scientists, the field needs further analysis of
the decisions that are made by crime scene investigators. The decisions
required to interpret evidence vary according to the issue being investigated, i.e., the question being asked. Although the crime scene investigator will commonly be expected to reconstruct the events (explain what
happened), on occasion he will be required to classify materials and to
individualize them. Consequently, courses must be designed to develop
facility in making all three types of decisions, not just how to properly
collect and document objects and the changes in them.
Crime Scene Investigation, Scientific Examination, Crime Scene
Science
B153 Assessment and In Vitro Repair
of Damaged DNA Templates
Ashley Hall, MS* and Jack Ballantyne, PhD, University of Central
Florida, 12354 Research Parkway Room 225, Orlando, FL 32826
After attending this presentation, attendees will have been provided
with methods for the assessment and repair of damaged DNA templates
derived from forensically relevant samples.
Little is known about the damage incurred to forensically relevant
DNA samples, and there are currently no methods for the repair of such
templates. We intend to present the results of our assessment studies, as
well as the sucessful repair of damaged DNA.
* Presenting Author
DNA extracted from biological stains is often intractable to
analysis. This may due to a number of factors including a low copy
number (LCN) of starting molecules, the presence of soluble inhibitors
or damaged DNA templates. Remedies may be available to the forensic
scientist to deal with LCN templates and soluble inhibitors but none
presently exist for damaged DNA. In fact, knowledge of the biochemical
nature and the extent of DNA damage in physiological stains is rudimentary at best. Also unknown is the point at which the damage inflicted
upon a particular sample precludes the ability to obtain a genetic profile
for purposes of identification. Therefore, the primary aims of this work
were first ascertain the types of DNA damage encountered in forensically relevant stains, correlating the occurrence this damage with the
partial or total loss of a genotype, and then to attempt the repair of the
damage by means of in vitro DNA repair systems.
The initial focus of the work was the detection of damage caused
by exogenous, environmental sources, primarily UV irradiation, but also
factors such as heat and humidity. By incorporating various lesion specific enzymes, a set of assays, both PCR and gel-based, have been
developed which describe the type and extent of damage inflicted upon
DNA, both in a hydrated and dehydrated state. By dividing the UV
spectrum into its component wavelengths, and combining each with
various other conditions, the major causes of damage have been identified and their effects on genetic profiling assessed.
Armed with this knowledge, the next focus was the repair of the
damage by means of in vitro DNA systems. Efforts have been concentrated on base excision repair, a direct reversal, single strand gap repair,
and translesion synthesis assays. By modifying the assays and
employing various combinations of the systems, a genetic profile has
been obtained from previously intractable samples.
DNA Damage, in vitro DNA Repair, UV Damage
B154 Studies of PCR Inhibition
Gary G. Shutler, PhD*; Tara L. Copp, BSc and Janice Chau, BS,
Washington State Patrol, Crime Laboratory Division, 2203 Airport
Way South, Suite 250, Seattle, WA 98134-2027
After attending this presentation, attendees will be able to identify
several problematic matrices for the DNA typing of bloodstains and to
evaluate the usefulness of a variety of techniques for alleviating PCR
inhibitory substances.
This information could be of assistance to crime labs to use as a reference for dealing with specific PCR inhibitor problems encountered
with STR analysis of casework material.
Bloodstains were prepared on 15 separate matrices; titanium, blue
denim jeans, lead, steel, drift wood, spruce 2x4, pressure treated spruce
2x4, maple tree bark, ocean beach sand, garden sand, compost soil, rock,
dried leaf, soiled dry leaf, and fresh grass clippings. DNA was prepared
from the bloodstains using a standard proteinase K digestion in buffer
and organic extraction followed by a Centricon ® spin procedure.
The most widely used method to avoid PCR inhibition is to use the
PCR facilitator, bovine serum albumin (BSA). Most forensic labs rely on
manufacturers’ kits for STR amplification. Both Applied Biosystems
(AB) and Promega have included BSA in their STR kit amplification
reactions. We decided to use the D1S80 AMFLP kit from AB (the last
few kits available from this discontinued product line), which does not
include BSA, as a model system to assess the effects of BSA on relieving
inhibition. DNA extracted from bloodstains on the 15 different matrices
was tested and examined for PCR inhibition. The DNA extracted from
bloodstains on titanium sheet metal, denim jeans, driftwood, 2x4 spruce,
99
maple tree bark, ocean beach sand, compost soil, rock, soiled dry maple
leaf and grass clippings exhibited D1S80 inhibition. The addition of
non-acetylated BSA (at 160µg/ml, New England BioLabs) relieved inhibition of DNA extracts from blood on the titanium sheet metal, the
denim, the rock, the dried maple leaf and the grass clippings. The
addition of acetylated BSA (Molecular Biology Grade, Sigma) did not
relieve inhibition. The results were then compared to results from the AB
AmoFISTR Profiler Plus™ kit. All DNA extracts from bloodstained
matrices that displayed inhibitory substances which could not be
relieved by BSA for the D1S80 also showed inhibited STR results.
There were 5 DNA extracts that did not show relief of PCR inhibition by BSA. These were driftwood, 2x4 spruce, maple tree bark,
ocean beach sand and compost rich soil. Several different techniques
were used in an attempt to reduce or alleviate the inhibition observed in
both the D1S80 and Profiler Plus results. These included extra
Centricon® or Microcon® spins, sepharose beads, chelex®, non-human
DNA on beads, QIAamp® Stool mini kit, DNeasy® Plant mini kit, diffusion in LMT agarose as well as other methods.
While some techniques were either effective or partially effective
for DNA extracted from any one particular matrix, there was no
universal solution for all matrices. A summary of the methods that
improved results for the problem matrices will be presented.
Data on the potential of DNA IQ to remove inhibitors from DNA
extracts and from bloodstains on inhibitory matrices will also be
presented.
Since there was no single solution, a test was devised to determine
if the inhibitory substance would be present in the final extract after
extracting an unstained matrix control. The extract was mixed with
control DNA and PCR inhibition was observed. The mixed inhibitory
extract-control DNA sample could be used to test for the most successful
method to relieve inhibition without wasting the limited critical evidence
sample. This approach could be useful in very special cases.
STRs, PCR Inhibitors, Bloodstains
B155 Case Studies: Interesting Kinship
Problems Solved During WTC
Identification Efforts
Noelle J. Umback, PhD*, Erik T. Bieschke, MS, Zoran M.
Budimlija, MD, PHD, and Sheila M. Estacio, MS, Office of Chief
Medical Examiner, Forensic Biology, 520 First Avenue, New York, NY
10016; Michael J. Hennessey, MBA, Gene Codes, Inc., 640 Avis Drive,
Ann Arbor, MI 48108; Felecia J. Lewis, BS, MPS, Elaine J. Mar, MS,
Carole Meyers, Bianca L. Nazzaruolo-Brandon, BS, MA, Anca M.
Nicholson, MS, and Mary-Breen O’Connor, BS, Office of Chief
Medical Examiner, Forensic Biology, 520 First Avenue, New York, NY
10016; Stephen F. Swinton, BS, MA, JD, and Peter Wistort, BS, New
York State Police Forensic Investigation Center, State Campus
Building #30, 1220 Washington Avenue, Albany, NY 12226; Robert C.
Shaler, PhD, Office of Chief Medical Examiner, Forensic Biology, 520
First Avenue, New York, NY 10016
After attending this presentation, forensic DNA analysts will be
informed of various types of pedigrees reconstructed, and other issues
faced, in order to identify victims from the WTC disaster.
Identification of remains from the WTC has brought closure to hundreds of victims’ families. Also, DNA and related information technology has been advanced far ahead of the normal pace of standard
research, in order to make those identifications.
Relatively easy DNA identifications can be made using known
victims’ blood samples (e.g. from the national bone marrow donor reg-
100
istry) or personal effects (toothbrushes, hairbrushes, razorblades, etc.),
supported by multiple familial DNA samples (parents and siblings,
spouse and children, or some combination thereof). The identification
process therefore also included the verification of not only all available
DNA data for all associated remains, but also the physical items and
information provided by victims’ families and friends, used in the DNA
identification process. Confirmation of who brought in what DNAbearing items for which victim is paramount. For instance, wives occasionally turned in their own toothbrushes instead of their husbands’;
well-meaning co-workers brought items in from workplace lockers of all
the missing persons from that location (at least one three-way switch is
known to have been made) or no collection information at all was
obtained (“donor name” was either left blank, or filled in with the
missing person’s name). Occasionally it was found that “fathers” or
“children” of victims really weren’t. All of these situations require substantial meta-data analysis prior to even attempting kinship calculations.
Kinship samples, used to validate personal samples of unknown or
unconfirmed origin (incomplete chains of custody were rampant), were
also used to make identifications outright. In addition, confirmed (either
by kinship analysis or by confirming the donor) personal effects in combination were used to make many identifications. One or more matching
personal effects in addition to at least one family member allowed much
higher confidence levels in those identifications.
Occasionally, analysts working on disaster identifications had to
“get creative” to solve intractable cases, especially ones where multiple
persons related either by blood or address (such as spouses or apartmentmates) both perished at the World Trade Center.
In one case, a brother and sister both died. She had no children, and
the personal item submitted for the sister yielded no DNA. However, the
brother had a wife and son; and one other sibling was available. Using
the sister, the missing brother’s comb, and the victim’s sister-in-law and
nephew enabled the identification of the missing sister in this pair.
The adult children of a couple who died in the tragedy, along with
her parents (his were not available) and his sister, submitted buccal
swabs for use in kinship analysis. A male fragment of remains came up
as a potential “hit” for this family. Using direct lineages from this
fragment to the couple’s children and his sister only, did not meet the statistical threshold set for this project, although it did indicate a strong
relationship probability. Addition of his in-laws’ DNA samples (as
grandparents of his children) to the pedigree made up for the fact his
wife’s DNA profile was unknown (her DNA, after all, had to have been
derived from her parents’), and raised the probability into the acceptable
range to declare the identification.
A pair of brothers were lost on 9/11. Both men’s wives donated personal effects in the form of razors and toothbrushes, and kinship swabs
were donated by their parents. In a case such as this, kinship alone is not
sufficient to show which son was which, since both are related in the
same way to their parents. One of the brothers was identified early on,
by kinship, before it was known that his brother was missing also. The
case records were reviewed and it was found that their files (and corresponding personal items) were jumbled together. The group of personal
effects had two male profiles between them which showed relatedness to
the parents of these two missing men. Eventually a distinguishing factor
was found between the two cases, and enabled analysts to determine
which profile came from which brother. Ultimately, remains were found
which matched both sets of items and both men were declared identified.
In general, the more samples which can be donated for a missing
person, the better. Problems ranging from poor DNA yields to wrong
DNA types can be often overcome by use of kinship samples to verify
that the submitted personal item was used by a member of a given
family, and that the profile fits well in the pedigree in the victim’s place.
Once a validated personal effect matches a disaster sample, that is an
identification.
World Trade Center, Kinship Analysis, Identification
* Presenting Author
B156 Case Study of ‘Deleted-Amelogenin’
Males Within the WTC DNA
Identification Project
Felecia J. Lewis, BS, MPS*, Noelle J. Umback, PhD, Erik T.
Bieschke, MS, Office of Chief Medical Examiner, Forensic Biology, 520
First Avenue, New York, NY 10016; Michael J. Hennessey, MBA, Gene
Codes, Inc., 640 Avis Drive, Suite 300, Ann Arbor, MI 48108; Elaine J.
Mar, MS, Anca M. Nicholson, MS, Lawrence Quarino, PhD, MaryBreen O’Connor, BS, Mechtild K. Prinz, PhD, and Robert C.
Shaler, PhD, Office of Chief Medical Examiner, Forensic Biology,
520 First Avenue, New York, NY 10016
After attending this presentation, attendees will be informed of the
“amelogenin deletion” phenomenon (known to occur in certain Asian
populations) which has been confirmed in the World Trade Center victim
population.
humanity by raising awareness in the forensic DNA community of
“amelogenin deletion,” and explain how this phenomenon was dealt
with during the WTC project in order to make a positive identification
of at least one victim.
Due to a known genetic mutation on the Y chromosome in which
the amelogenin locus is deleted (1, 2), STR-typing kits such as Cofiler,
Profiler Plus, and PowerPlex™ 16 only detect the copy of the allele
which originates on the X chromosome. As a result, the genotypes
appear to be female, unless shown to be otherwise by use of Y-STR
typing. This rare “Y-deletion” is found most often in males from the
areas between Southeast Asia and the Indian subcontinent.
The World Trade Center disaster included many persons whose
ancestry can be traced to that region of the world, and the presence of Ychromosome anomaly has been confirmed during the identification
process for one victim, and is suspected in another. The latter was identified by dental records (his body was intact enough at recovery to still
be wearing masculine clothing, but gave a “female” profile in autosomal
STRs. Thus, for the ~ 2800 reported missing (about three quarters of
which are male), the prospect of observing this twice is somewhat
remarkable.
During DNA testing for the World Trade Center identification
project, two human remains fragments (both are pieces of bones) found
at the site matched a personal effect submitted by a male victim’s family.
The match was initially thought to be a “problem case” such as comingling (two persons’ remains, and therefore DNA as well) or a sample
mixup (for example, another family member’s toothbrush submitted by
mistake as an exemplar for their missing loved one), because the personal sample for this missing man came up with a female profile during
initial testing (Cofiler and Profiler Plus). In addition, several members of
his family submitted buccal swabs for use in kinship analysis, including
his father and a full brother. It was found that all of this family’s samples
appeared to be female, but in all other loci tested, the correct allele inheritance patterns indicated that there likely wasn’t a mixup of the swabs.
These results bolstered our suspicion that the Y chromosome in that
paternal line was “deleted-amelogenin.”
Additional testing using Y-STRs (OCME’s YM1 panel, consisting
of DYS 19, DYS 389 I and II, DYS 390) confirmed an identical Y haplotype in the father’s and brother’s buccal swabs and the victim’s toothbrush. That result confirmed our suspicion of the Y-chromosome
mutation’s presence in this family; and taken together, all results were
enough to declare a positive DNA identification of this WTC victim.
Laboratories performing DNA typing involving victims, suspects,
or missing persons (specifically, males) from southern and eastern Asia
must be aware of the possibility of a situation like the above arising, not
only in identifications when kinship samples are also available to
confirm the phenomenon, but also during criminal casework such as
homicides and sexual assaults when an unexpected additional female
* Presenting Author
profile is obtained. Additional, Y-chromosome-specific, testing can be
performed to help solve these cases.
1. Santos FR, Pandya A, Tyler-Smith C. Reliability of DNA based sex
tests. Nat. Genet. 1998; 18:103.
2. Roffey PE, Eckhoff CI, Kuhl JL. A rare mutation in the amelogenin
gene and its potential investigative ramnifications. J. Forensic Sci.
2000; 45:1016-1019.
Y-Deletion, World Trade Center, Amelogenin
B157 Operation Iraqi Freedom: DNA
Identification, Confirmation &
Reassociation of Combat Casualties
Based Upon Reference DNA Samples
David A. Boyer, MFS*, Deprtment of Defense DNA Registry, 16050
Industrial Drive, Suite 100, Gaithersburg, MD 20877; Demris A. Lee,
MSFS, Armed Forces DNA Identification Laboratory, 1413 Research
Boulevard, Rockville, MD 20850; Brion C. Smith, DDS, Department of
Defense DNA Registry, 1413 Research Blvd, Rockville, MD 20850;
Lisa Pearse, MD, MPH, Armed Forces Medical Examiner System,
1413 Research Boulevard, Rockville, MD 20850; Russell Strasser, MFS
and Craig T. Mallak, MD, Armed Forces Medical Examiner System,
1413 Research Boulevard, Rockville, MD 20850; Garry J. Alexander,
BS, Dover Port Mortuary, 121 Carson Drive, Dover AFB, DE 19902
After attending this presentation, attendees will be provided with
details of the Armed Forces DNA specimen collection program and its
utility in providing ready references for issues of human identification.
humanity by facilitating awareness of the benefits of a DNA reference
database to successfully identify military war casualties.
This presentation will detail the casualty identification process of
Operation Iraqi Freedom (OIF), report the various identification
methods, and highlight the role of the U.S. Department of Defense DNA
reference sample repository facilitating that process.
authors and are not to be construed as official or as the views of the U.S.
Department of Defense or the U.S. Department of the Army.
The war against Iraq began on March 20, 2003 and officially ended
on May 1, 2003 with the U.S. military gaining control over the capitol
city of Baghdad. Although hostilities continued to produce casualties
this discussion is limited to the initial combat action. The 43 day war
with Iraq resulted in 142 deaths of Army, Air Force, Marine, Navy, and
U.S. civilian personnel. These casualties were recovered from the battlefields, prepared for shipment in country at the Theater Mortuary
Evacuation Point, and transported to the Port Mortuary, Dover Air Force
Base, DE, for identification. Two of the casualties were sent to medical
facilities in Europe rather than Dover, DE. The 140 sets of remains
arriving at Dover were examined by teams of forensic pathologists,
odontologists, anthropologists, and fingerprint experts for identification.
Every suitable piece of recovered human remains was sampled for DNA
testing either as the primary method of identification, a confirmatory
means to supplement other identification methods, or for reassociation
of fragmented remains.
Operation Iraqi Freedom and Operation Enduring Freedom in
Afghanistan marked the first war efforts undertaken by the U.S. Forces
since the establishment of the Armed Forces Repository of Specimen
Samples for the Identification of Remains (AFRSSIR) in 1991. The
repository is a warehouse of dried bloodstain cards used as ready DNA
references in human identification of service members. All military personnel and select civilian employees working for the military are
required to have a DNA specimen on file at the AFRSSIR. The
AFRSSIR is the largest DNA reference database in the world and contains more than 4,000,000 DNA reference samples. It has been used
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more than 1,000 times in military identifications since 1995. The references are retrieved as required from cold storage in Gaithersburg, MD
and transported to the Armed Forces DNA Identification Laboratory in
nearby Rockville, MD for analysis. Results are used for comparison to
biological specimens obtained from the remains at autopsy.
Combat deaths in OIF ranged from single gunshot wounds to
massive explosions. Deaths resulted from hostile actions, homicides,
accidents, suicides, and natural causes. Some deaths were classified as
non-hostile, a casualty due to circumstances not directly attributable to
hostile action or terrorist activity. The condition of OIF casualties ranged
from fully intact bodies to highly fragmented remains, some exhibiting
varying stages of decomposition.
There were 135 AFRSSIR record specimens on file to aid in the
identification process. Four military casualties and three U.S. civilians
were not represented in the AFRSSIR database. Of the 140 casualties
processed at the Dover Port Mortuary, 89 were identified by a combination of fingerprint examinations, dental comparisons and DNA testing.
Eighteen identifications were made by fingerprints and DNA, ten were
identified by fingerprints and dental examination, and six were identified
by dental exam and DNA testing. There were 13 identifications based
solely on DNA testing, two on fingerprints only, and two independent
identifications from dental examination. DNA analysis successfully
reassociated 129 fragmented remains to 25 casualties. Five of the human
remains sampled for DNA yielded no results or insufficient data to
render a conclusion. Three DNA profiles did not match reference
samples for any of the U.S. casualties. There were a total of 103 other
fragments returned from Iraq that were unsuitable for DNA testing,
deemed either inappropriate material or classified as non-human
remains.
None of the U.S. Forces who died in the 43 day battle of Operation
Iraqi Freedom went unidentified. The military’s mandated DNA reference specimen collection program was highly successful in providing
direct references of reported casualties for DNA analysis. The AFRSSIR
DNA references facilitated rapid testing which yielded full DNA profiles
and resulted in expeditious war casualty identification, identification
confirmation and reassociation.
Operation Iraqi Freedom, DNA Repository, DNA Identification
B158 A Comparison of Automated
DNA Extraction Methods
Daniel P. Cheswick, BS*, Mechthild Prinz, PhD and Robert Shaler,
PhD, NYC Office of the Chief Medical Examiners, 520 First Avenue,
New York, NY 10016
This presentation will describe and compare three different automated DNA extraction approaches.
Three automated extraction procedures were evaluated and compared to chelex extraction. The current chelex extraction procedure used
at the OCME takes approximately 2 hours to complete 20 samples. If
this step could be automated, it would free up the analyst to work on
additional tests.
The comparison experiments were designed to address the success
rate for small stain sizes and the efficiency of the removal of inhibitors.
Bloodstains of 1 µl and 4 µl in size were made on white and dyed cotton
cloth, filter paper, suede, blue jeans, and rug. The stains were then
extracted using 5% chelex (OCME STR manual, 2003), QIAamp™ by
Qiagen, DNA IQ™ by Promega Corporation, and MagAttrct™ by
Qiagen.
QIAamp kits automate the isolation of nucleic acids from a wide
variety of samples with a 96-well plate procedure. Nucleic acids bind
specifically to the QIAamp silica-gel membrane while contaminants
pass through. While the extraction yields a large amount of purified
DNA, the batch size is not flexible. The QIAamp technology is currently
running on a Biorobot 9604.
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MagAttract DNA Systems are designed for fully automated DNA
purification on a M48 or a M96 robot. The M48 has many anti-contamination features such as an enclosed workstation, a drip tray, and built in
UV capability. MagAttract uses paramagnetic particle technology for the
capture and release of DNA. The batch sizes of the M48 and M96 robots
are flexible.
The DNA IQ™ system is a DNA isolation system and quantitation
system designed to yield a preset constant amount of DNA. This constant amount of DNA allows quantitation to become optional on
database samples. This system also employs aparamagnetic particle
technology that binds and releases the DNA. Excess DNA is removed
during the wash steps because of the paramagnetic particles becoming
over saturated. The isolation of genomic DNA using the DNA IQ™
System has been automated on the Biomek™ 2000 Laboratory
Automation Workstation.
All extracted samples were quantitated using the Quantiblot™
system by Perkin Elmer Applied Biosystems. Selected samples were
then amplified using Profiler plus with the product then ran on a ABI
3100 DNA Sequencer.
Data were evaluated based on the amount of DNA recovered,
presence or absence of inhibitors, and amplification success rates.
Automation, STR Testing, Extraction
B159 High Throughput STR Testing of Tens
of Thousands of 9- 12-Year-Old Bone
Samples From the Former Yugoslavia
Using a Silica Based Extraction Method
Jon M. Davoren, MS*, Daniel B. Vanek, PhD, Rijad Konjhodzic, MS,
Scott Strong, BS, and Edwin F. Huffine, MS, International Commission
on Missing Persons, Alipasina 45a, Sarajevo, 71000, Bosnia and
Herzegovina
Attendees will be presented with a review of the International
Commission on Missing Persons high throughput bone STR testing
facility including optimization of extraction, amplification and the minimization of costs.
The results presented would have benefit to anyone performing
STR analysis of difficult samples such as bone. These results may also
be of benefit to other forensic projects whose goals are also to use DNA
STR tests in order to confirm the identify of mortal remains. These
results were obtained from possibly the largest mass identification
process ever attempted.
The ICMP has developed a DNA-led identification effort to assist
in the identification of the estimated 30,000 – 40,000 persons that went
missing during the breakup of the former Yugoslavia. Due to the large
number of missing persons and the lack of medical and dental records
for this region the strongest piece of evidence that will lead to identification is a DNA STR profile. In the past large scale bone STR testing
has not been feasible because of numerous limitations. Some of the limitations to performing large scale DNA testing on bone samples are the
high costs, the high rate of failure in the testing process, the large
number of loci needed to generate significant results, and the time
required for such testing.
To increase the success rate of STR testing on bone samples a
silica-based extraction method was developed and has been shown to be
much more successful than that of the organic extraction method. To
directly compare the DNA isolated by these two methods real time PCR
was performed using the Applied Biosystems Quantifiler® kit on the
ABI 7700 Sequence detection system. Preliminary results of this comparison show that the ICMP developed silica-based extraction protocol
both isolates more DNA than the organic method as well as it reduces the
levels of PCR inhibition that are often observed during amplification of
DNA from bone samples. Further studies using the Quantifiler kit have
* Presenting Author
allowed the optimization of PowerPlex® 16 and SeFiler® amplification
reactions based on the estimated DNA content recovered from bone
samples.
Using this silica based extraction system in combination with the
Promega Power Plex® 16 system the ICMP has extracted nearly 22,000
bone samples for around 11,000 cases. Using this system at least 14 loci
have been amplified for approximately 10,000 of these cases. For some
cases where individual loci failed, the initial testing, attempts were made
to amplify these loci using a single primer pair as a monoplex system has
some advantages over the 16-plex system.
The average consumable reagent costs per case of bone STR
analysis including all consumables is less than $100USD for duplicate
bone sample extractions and $85USD for extraction of two teeth. This
processing system in combination with the relatively inexpensive cost of
labor in the former Yugoslavia has led to a rapid, relatively inexpensive
system that is currently identifying 300 – 400 bodies per month.
DNA, STR, Real Time PCR
B160 Recovering Biological Samples From
Crime Scene Using FTA Paper
Jose A. Lorente, MD, PhD*, Luis J. Martinez-Gonzalez, MS,
Francisco Fernandez-Rosado, MS, Esther Martinez-Espin, MS, J.
Carlo Alvarez, PhD, Carmen Entrala, PhD, Miguel Lorente, MD, PhD
and Enrique Villanueva, MD, PhD, University of Granada,
Department of Legal Medicine, Faculty of Medicine, Granada, 18012,
Spain
Attendees will learn to recover biological specimens from crime
scenes using a novel approach that could facilitate the future practical
work in the future.
This is a practical approach to help in one of the main problems in
crime scene investigation and in DNA labs: how to properly recover,
store, and analyze biological samples.
Proper collection and preservation of biological evidences
recovered from the scene of a crime is crucial to facilitate the analysis
and interpretation of all analytical results, including DNA typing.
FTA® paper is a well-known and widely used media to collect and
store biological materials for DNA or RNA analysis. Its uses are primarily focused on the generation of clinical and forensic databases,
although it has also been used for preservation of other biological materials, such as plant, animal, and bacterial specimens.
One of the main advantages of FTA is its ability to preserve biological materials at room temperature. When specimens are spotted or
applied to FTA cards, cell membranes and organelles are lysed, and the
nucleic acids are released, causing both DNA and RNA to become
entrapped in the fibers of the matrix. FTA cards rapidly inactivate
organisms, including blood borne pathogens, and prevent the growth of
bacteria and other microorganisms. Therefore, biological samples such
as blood or saliva can be preserved at room temperature. It is also
important to mention that archived samples are ready for analysis in less
than 30 minutes.
Because of the above advantages, we are evaluating the utility of
FTA paper as a reliable support to collect and store biological specimens
that are usually found and collected at the scene of crime. For this study
we have spotted blood samples (1, 5, and 10 microliters) on different
surfaces (wood, cotton, tile, glass, and carpet); samples were collected
after 3 different periods of time: one day, one week, and one month. Two
different methods of recovery are being evaluated. The first method is
moistening the FTA paper with sterile distilled water and then pressing
the FTA paper on the stain; the second method is moistening the dried
stain with sterile distilled water and then blotting it with the FTA paper.
DNA purification was performed following the recommendations
* Presenting Author
from the manufacturer (Whatman Inc., Clifton, NJ), using FTA purification reagent and 1.2 mm punches. DNA analysis was performed using
both PowerPlex16 (Promega), and Identifiler (Applied Biosystems).
Results were visualized in a capillary electrophoresis system, model
ABI-310 (Applied Biosystems).
Results show that FTA paper is an ideal medium to collect dry
blood specimens from hard, non-abosrbent surfaces, such as glass, tile,
and wood, where it has been possible to recover and amplify DNA from
very small samples of blood, as small as 1 microliter. It’s also a good
media to collect samples from absorbent surfaces such as carpet and
cotton, although the performance depends on the size of the stain
(amount of microliters of blood spotted). The best approach in our hands
is to first moisten the evidence, and then apply and press with FTA paper.
We are currently evaluating the usefulness of FTA paper to recover
saliva and semen samples from mock crime scenes, with positive preliminary results.
Considering the above mentioned results, FTA offers several
advantages as a support for collecting and preserving biological samples
recovered from the scene of crime. First, it is simple to recover samples
from the crime scene since it is only necessary to wet and apply the FTA
paper. Second, it is possible to transport and store samples at room temperature for a long time. Third, the original substrate (the place where
the blood was deposited in the crime scene) is preserved, since there is
only a transfer of cells from the substrate (i.e., the wood or the tile) onto
the FTA matrix. Finally, a fourth advantage to consider is the use of a
single method for collecting and storing different kind of samples that
would be achieved by using FTA for crime scene collection.
FTA, Evidence Collection, DNA Analyisis
B161 DNA Analysis of Digested
Seeds in Forensic Samples
Cheng-Lung Lee, MFS*, National Tsing Hua University and Hsinchu
Municipal Police Bureau, 1 Chung Shan Road, Taiwan 300, ROC;
Heather Miller Coyle, PhD, Eric Carita, MS, Carll Ladd, PhD,
Nicholas C.S. Lang, MFS, and Timothy M. Palmbach, Esq, Department
of Public Safety, Division of Scientific Services, 278 Colony Street,
Meriden, CT 06451; Ian C. Hsu, PhD, Nuclear Science Dept, National
Tsing Hua University, 101, Section 2 Kuang Fu Road, Hsinchu, Taiwan
300, ROC; and Henry C. Lee, PhD, Forensic Science Department,
University of New Haven, 300 Orange Avenue, West Haven, CT 06516
Seed and other plant materials are valuable evidence not only for
linking a suspect to a particular crime, but also important for the verification of an alibi, or to provide new investigative leads during criminal
investigations. Currently, microscopic examination of the materials in
vomit, stomach content, or feces is the only method of choice for the
identification and association of seeds to possible sources.
In recent years, DNA has been successfully extracted and analyzed
from a variety of plant materials. Many cases have been solved due to
the DNA linkage of plant materials to their source. However, in
reviewing the literature, very few studies have been conducted on the
subject of recovery and analysis of seed DNA.
Our research results showed that high quality DNA could be
extracted from a variety of seeds after they went through the human
digestive system. DNA also was successfully recovered from seeds in
human feces. A comparison of DNA yield on two types of extraction
techniques (Hand grinding, Mechanical crushing-Mixer Mill method)
will also be reported. Our preliminary data on DNA analysis with the
AFLP technique on 5 types of tomato seeds and 4 types of pepper seeds
indicates that DNA-AFLP is a viable procedure for the identification and
individualization of seeds in forensic investigation.
Forensic Botany, Seed Evidence, AFLP
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B162 Sampling Considerations in the Analysis
of Glass Fragments by Laser Ablation
Inductively Coupled Plasma Mass
Spectrometry (LA-ICP-MS)
Tatiana Trejos, BSc* and José R. Almirall, PhD, Florida International
University, International Forensic Research Institute, 11200 SW 8th
Street, CP 194, Miami, FL 33199
After attending this presentation, attendees will have an understanding of the advantages of the new technique of laser ablation ICPMS for the elemental analysis of glass fragments with a focus in the sampling strategies that should be taken into account during analysis.
The presentation will describe the advantages of the laser ablation
sampling for ICP-MS analysis of glass evidence, including fast analysis
time, excellent discrimination power and minimum consumption of
sample. The presentation intends to give continuity to the results previously presented at AAFS. Different studies of interest to the forensic
practitioner will be presented including sampling strategies and considerations for glass analysis by LA-ICP-MS.
The authors have previously reported the use of Laser Ablation
ICP-MS as a powerful analytical tool for elemental analysis of glass.
The advantages of LA-ICP-MS include fast, sensitive determinations of
the trace metals in glass with a minimum destruction of the evidence.
Glass fragments recovered from crime scenes are typically of 0.1 to
1mm in length and therefore in some cases the amount of sample is very
limited to perform conventional digestion methods, which consume at
least 2-3 mg of glass per replicate. LA-ICP-MS requires ten thousand
times less glass consumption per analysis (~250 ng) and therefore the
sample remains practically unaltered. However, due to the minute
amount of material removed in LA, the analyst should be aware of
special sampling considerations such as characterization of the glass
fragments originating from the “known” source, fragment size and
selection of the area and surface of ablation.
The fact that glass fragments collected from the crime scene and
from suspect(s) and victim(s) are random in size does not affect bulk
digestion analysis because sample is crushed and homogenized before it
is weighed. During laser ablation, small craters (~50µm) are drilled into
the solid material and therefore an investigation of the effect of the size
of the fragment on the quantification of glass was conducted in order to
determine if the interaction of the laser-surface changes significantly
with the size of glass. Standard reference materials SRM 612 and SRM
610 were selected to conduct this part of the work in order to account for
different concentration ranges and different opacity of the samples. The
set under study was comprised of 7 fragments originating from each
standard at different sizes and shapes ranging from 6 mm to 0.2 mm in
length. The results show that there is not a significant difference in the
elemental composition of fragments of different sizes. In addition, a
homogeneity study of the elemental composition of glass samples was
performed on glass matrices commonly found in crime scenes such as
containers, architectural windows, windshields and headlamps. The set
of glasses under study was comprised of 56 samples originating from
glass containers, 28 samples from automobile windshields, 20 samples
from architectural windowpanes and 20 samples from vehicle headlamps. All fragments were selected with a size smaller than 2 mm in
length in order to simulate the typical glass fragments transferred from
the crime scenarios. A CETAC LSX 500 Nd:YAG laser, 266nm, flat top
beam profile was used in single point mode sampling 50 µm spot size
for 50 seconds at 10 Hz (500 shots). The isotope 29Si was used as an
internal standard and the standard reference material, SRM NIST 612,
was used as a single point for the external calibration. The standard SRM
621 was used as another control for the containers set, SRM 1411 for
headlamps and SRM 1831 for the automobile and architectural window
sets due to their very similar matrix with the samples of interest. For
104
each set of glasses, the mean values and standard deviations of ten replicates (n=10) of a single fragment were compared with the values
obtained from ten (n=10) different fragments of glass within the area of
interest in order to evaluate whether or not the variation within a glass
was larger than the variation due to the method. An additional subset of
tempered glasses was examined to perform an elemental composition
profile within different depths of the fragments. Single shot (one laser
pulse per analysis) was also evaluated and its limitations for the forensic
analysis of glass are also presented. The method presented for the
analysis of glass by LA-ICP-MS had previously shown to possess the
same or better performance than dissolution –ICP-MS methods in terms
of accuracy, precision, limits of detection and discrimination power. The
results show that glass is homogenous even at the micro-range level
allowing LA-ICP-MS as an alternative technique to perform elemental
analysis of glass. However, the variation of elemental composition of
headlamps and containers is larger over the source than the instrumental
variation due to inherent heterogeneity and therefore different statistical
tools are recommended to fully characterize glass originating from the
known source before the comparison analysis can be made. The proposed method allows reliable forensic comparisons of glass fragments
by LA-ICP-MS independently of the fragment size recovered at scene.
Glass, Laser Ablation ICP-MS, Elemental Analysis
B163 Elemental Analysis by LA-ICP-MS
at the Netherlands Forensic Institute
Shirly Montero, PhD*, Maarten Hordijk, Ir, Wim Wiarda, Ing, Peter de
Joode, Ing, and Gerard J.Q. van der Peijl, PhD, Netherlands Forensic
Institute, PO Box 3110, Rijswijk, ZH 2280 GC, Netherlands
After attending this presentation, attendees will understand the discrimination potential of LA-ICP-MS for the elemental analysis of different types of materials of forensic interest.
This presentation will encourage the adoption of methods with the
high discrimination potential for forensic applications rendered by the
(LA-)ICPMS technique. This presentation focusses on a few of these
applications.
The value of trace evidence has been established for some time
through numerous studies and publications. Recovered samples associated with a suspect and samples associated with a known source sometimes may be compared to each other based on the optical and other
physical properties as well as the chemical composition of such samples.
Moreover, such comparisons have been shown to be useful as a source
of investigative information, not only for the associative purpose mentioned above such as relating a suspect to a crime scene but also for
reconstruction of events. The significance of the results of these comparisons can only be estimated by the analysis of the information
obtained from appropriate databases. The choice of methods to characterize and compare trace evidence depends on, among other factors, the
size of such evidence. It is known, for example, when a piece of a
material is broken, the fracture surface will often have unique topological features. When the surface is large enough, the features produced
by the fracture may be so abundant as to permit a distinctive association
between two sections, like in two pieces of a jigsaw puzzle. This
physical matching is the most definitive way of relating samples that
were once a single piece. However, when the physical matching is not
possible other properties can be measured.
The decrease in the variation of physical properties in evidence of
the same kind, i.e. by increased quality control by manufacturers, limits
the discrimination potential of the methods used to measure such properties. The use and value of elemental analysis for characterization of
materials of forensic interest becomes important in such cases.
Furthermore, when the dimensions of a sample are small, very sensitive
methods are needed in order to measure reliably the elemental composition, in particular at trace levels. The last few years, efforts have been
* Presenting Author
made by different research laboratories to develop and validate analytical methods with better sensitivity, precision and reproducibility.
Various instrumentation available in forensic laboratories has been
used for the elemental analysis of evidence samples (e.g., glass, paints,
fibers), among them SEM-EDS, XRF, ICP-AES, ICP-MS and more
recently LA-ICP-MS. This last technique combines the sensitivity and
precision of ICP-MS with the advantages of laser ablation sampling.
With the use of LA-ICP-MS there is no need for laborious and lengthy
digestion procedures with dangerous chemicals. In addition, the
common analytical interferences that are increased by the presence of
solvents are minimised using laser sampling, improving the detection
limits of some potentially discriminating elements. The amount of
material ablated for the complete analysis is very small (~fg), allowing
the analysis (including replicates) of very small samples. The destruction
is minimal with craters in the order of 10-8 m2, in contrast to the solution
approach where the whole sample to be analysed is irreversibly digested.
For this study, analytical methods for the elemental analysis by LAICP-MS of materials of forensic were optimised and validated in our laboratory at the Netherlands Forensic Institute. The system used is an ICPMS (Perkin Elmer ELAN6100 DRC) in combination with a 3 mJ- 213
nm-laser ablation system (New Wave UP-213). The standard reference
materials (SRMs) NIST 612, 1830 and 1831 were used for the development and validation of a quantitative method for glass analysis. The
SRM NIST 612 is a synthetic glass of well-characterised trace elemental
composition. This SRM was used to select an appropriate elemental
menu based on reproducibility and accuracy. The other two SRMs are
not certified for all trace elements but they were used for the optimization of the method for analysis of float glass (similar matrix) as well
as controls to monitor the stability and performance of the instrument
during the analysis. This method was then used to analyse ~200 float
glass samples collected by the police from different locations within the
Netherlands where a burglary had taken place. Although these samples
were not submitted as evidence they are considered representative of
possible casework samples. In addition, the refractive index of all the
samples was measured using a GRIM II. Less sensitive semi-quantitative elemental composition measurements were performed using XRF.
All samples were inter-compared using elemental compositions and
refractive index values. These comparisons were performed using the
SPSS statistical software package and were used to evaluate the discriminating value of each element measured with the two methods in
addition to that for the refractive index. Both the methods and the results
of the comparisons of the samples will be discussed in this presentation.
In addition, the development and validation of semi quantitative
methods for the analysis of other materials of forensic interest such as
automotive paints, inkjet inks and amphetamines will also be addressed.
This paper will evaluate laser ablation-inductively coupled plasmamass spectrometry (LA-ICP- MS) as a technique to determine the
unique trace elemental signature or fingerprint of glass samples for discrimination, and present criteria and protocols for the comparison and
differentiation of glass fragments from different sources by LA-ICP-MS.
Characterization of glass fragments is normally accomplished by
measuring the physical and optical properties of density and refractive
index. However further discrimination, such as identification of a suspected source of origin, has become more difficult as the range of
refractive indices has narrowed within glass subtypes because of
advances in glass manufacturing technology.
LA-ICP-MS provides a means for the forensic examiner to assign a
probability for positive association of a questioned glass sample based
on its trace elemental composition. Knowing trace element concentrations improves the confidence of a match and strengthens the value of
evidence presented in court. ICP-MS provides a high level of discrimination for glass samples due to excellent detection limits (10-100 times
better than ICP-atomic emission spectroscopy), wide element range coverage, and isotopic information. LA-ICP-MS is rapid, eliminates the
need for extensive sample preparation, and is virtually a nondestructive
technique, allowing for the possibility of further analysis of questioned
samples by corroborative techniques. Furthermore, LA promises to
increase the number of analytically useful elements by eliminating
problems with some elements due to poor dissolution and contamination. Additionally, smaller samples may be analyzed making the technique applicable to more cases.
This study focuses only on standard residential window and tempered glass, indistinguishable by refractive index measurements, provided by the Illinois State Police Forensic Sciences Command. After the
elemental fingerprints (i.e., mass spectra) and concentrations were
acquired for the provided samples, the criteria for comparison and differentiation were determined. The primary approaches evaluated for
classification/differentiation were: the presence or absence of elements;
comparison of the abundance of selected elements; comparison of relative abundance of elements for specific association patterns; and multivariate analysis of the complete mass spectrum.
The multivariate analysis technique used, was Principal
Components Analysis (PCA), which is an analysis tool for data compression and information extraction. PCA provides rapid analysis of
samples without time-consuming pair-wise comparison of calibrated
analyses. In general, PCA reduces a set of data into its most common
variables or factors and expresses these variables as eigenvalues or
scores that describe the major trends and variations in the data set. The
scores can then be graphically used for discrimination of glass samples,
since they provide an accurate description of the entire data set.
LA-ICP-MS, Elemental Analysis, Trace
Glass Analysis, LA-ICP-MS, Chemometrics
B164 Laser Ablation Inductively Coupled
Plasma Mass Spectrometry of
Forensic Glass Samples
David Baldwin, PhD, R.S. Houk, PhD, and David Aeschliman, PhD,
Ames Laboratory, B5 Spedding Hall, Iowa State University, Ames, IA
50011; Stan Bajic, PhD*, Ames Laboratory, 144 Spedding Hall, Ames,
IA 50011
After attending this presentation, attendees will have been presented with laser ablation-inductively coupled plasma-mass spectrometry as a technique to differentiate glass samples with similar optical
and physical properties.
The results from this study will demonstrate that LA-ICP-MS is a
fast, accurate, and reliable technique that provides a high level of discrimination in the analysis of glass fragments that cannot be differentiated by other techniques.
* Presenting Author
B165 Interlaboratory Validation Studies of
ICP-AES and ICP-MS Methods for
Elemental Analysis of Bullet Lead Alloys
Robert D. Koons, PhD* and JoAnn Buscaglia, PhD, FBI Laboratory,
CTFSRU, Building 12, FBI Academy, Quantico, VA 22135
Attendees of this presentation will learn the relative merits and limitations of using ICP-AES and ICP-MS methods in a forensic laboratory
for the sourcing of bullet lead.
Practical information which will assist forensic laboratories in
determining the feasibility of utilizing ICP-AES and ICP-MS for elemental analysis of bullet lead in their laboratories; results of interlaboratory validation studies will help support the legal admissibility of these
methods for bullet lead comparisons.
Elemental composition of the lead portion of bullets and shot
pellets may be used by forensic laboratories for comparison of two projectiles to see whether they may have a single production source. Of
105
more recent concern is the ability to determine a source, such as country
of manufacture, of one or more bullets used in a terrorist event. In an
international cooperative effort, NITE-CRIME Network has developed
analytical protocols for the use of natural isotopes and trace elements in
a variety of materials, including bullet lead. As part of this task, consideration was given to the fact that, while ICP-AES has long been used for
the comparison of bullets, quadrupole ICP-MS instruments are seeing
growing popularity and may be better suited to projectile sourcing.
Interlaboratory studies were conducted using both ICP-AES and ICPMS to evaluate the performance of each method for this purpose. The
results of studies considering the relative merits of combinations of each
instrument with dissolution protocols, both with and without lead
removal, and direct solid sampling by laser ablation will be presented.
Figures of merit, such as precision and accuracy, are related to the
specifics of sample digestion and instrumental operating parameters.
Specific recommendations will be made concerning some practical considerations in the utilization of these instrumental methods in the
forensic laboratory.
Bullet Lead, Elemental Analysis, Interlaboratory Study
B166 IRMS and ICPMS Studies
on Packaging Tapes
Gerard J.Q. van der Peijl, PhD*, Andrew M. Dobney, PhD, and Wim
Wiarda, Ing, Netherlands Forensic Institute of the Netherlands
Ministry of Justice, PO Box 3110, Rijswijk, 2280 GC, Netherlands
After attending this presentation, the participant will appreciate the
strong potential of the combination of ICPMS and IRMS techniques, not
only for tape investigations but for a wide spectrum of forensic investigations.
This presentation will demonstrate new interesting forensic applications of the (LA-)ICPMS and IRMS elemental and isotopic techniques
presently developed. They promise to result in much more strongly discriminating methods for forensic applications. The subject in the present
presentation is just one of these applications but is very useful in demonstrating the relevance of these techniques. The relevance is also recognized by the two international forensic networks that have become active
in this field: NITECRIME and FIRMS. More information on these networks can be submitted if requested.
Results are presented for tape comparison experiments using the
novel forensic IRMS (Isotope Ratio Mass Spectrometry) technique.
Results will be discussed and compared with results using other new
forensic techniques such as solution nebulisation ICPMS and LAICPMS.
Introduction: Brown packaging tape is encountered in 60% of the
violent crimes (murder, rape) committed in The Netherlands where tape
is found at the crime scene. Normally, the forensic scientist is asked to
compare the tape retrieved from the crime scene with tape found with a
suspect. At the NFI a combination of visual investigation (physical fit,
tape dimensions, colour, morphology), FT-IR and XRF is used routinely
to compare tapes. FT-IR can be used to identify the type of glue and
backing polymer. A combination of visual comparison and XRF analysis
generally suffices to discriminate between different tape products but
cannot be used for further discrimination between different batches of
one brand of tape product.
Solution: ICPMS In earlier studies, the more varying adhesive
layers of the tapes were investigated using both solution nebulisation
ICPMS (Inductively Coupled Plasma Mass Spectrometry) and LAICPMS. With these techniques trace levels of elements are detected and
identified. Solution nebulisation ICPMS experiments, especially, produced very discriminatory results. A ThermoFinnigan HR-ICPMS was
used. For solution nebulization measurements, the glue (ca 50 mg) was
first separated from the backing material through mobilisation in a
solvent (methanol, hexane) and then digested in a microwave oven (75
106
bar, 290 ºC) in a nitric acid/H2O2/H2O mixture. For the three different
brands investigated, commercial samples from the same tape product
acquired at different times from one commerical outlet and at one time
from different outlets could be discriminated on the basis of the solution
nebulisation ICPMS results whereas visual comparison and XRF (macro
elements) were not sufficient for discrimination between these tapes.
LA ICPMS Laser ablation measurements on intact tape samples of
the above rolls were made directly on the glue layer using a 266 nm
Nd:YAG laser. Tape samples were placed in the sample chamber and the
glue layer was laser ablated. Volatiles and aerosols produced in this way
were swept into the ICPMS system (low mass resolution mode). As
optimal values a laser pulse energy of 2 mJ, a pulse repetition rate of 10
Hz, a spot size of 80 µm and an ablation distance (lateral shift of laser
spot) of 30 µm were chosen. Laser spots were therefore partially (50 µm)
overlapping. Signals were integrated for 60 seconds over a grid area of
1 mm2.
LA ICPMS results for the different rolls of one brand of tape
demonstrate for each brand that discrimination power is sufficient to discriminate the rolls but is somewhat lower relative to solution nebulization results. In these experiments all tapes could be discriminated.
Upon repetition of the LA ICPMS experiments for the three different
rolls of one brand on another day the same distribution pattern is
observed. The exact location of the distribution pattern is not exactly the
same however, reflecting in our opinion variations in laser pulse energy
observed during these specific experiments.
IRMS (Isotope Ratio Mass Spectrometry) focuses on stable isotope
ratios of abundant elements in the samples such as H, C and O (O only
in the oxygen containing materials). Isotope ratios used were 2H/1H,
13C/12C and 18O/16O. Tape samples were analysed for us at IsoAnalytical Ltd (Sandbach, Cheshire CW11 3HT, UK). The IRMS used
was a Europa Scientific Geo 20-20 instrument. All samples were measured in triplicate. Tape samples were prepared for analysis by separating the glue and backing layers which were analysed separately. Also
complete tape samples, without further sample preparation, were
analysed.
IRMS Conditions: Hydrogen isotope analysis (ca. 6 mm2 tape
sample) was conducted by total conversion at 1080 ºC in a quartz reactor
lined with a glassy carbon film, filled to a height of 180 mm with glassy
carbon chips. Hydrogen was separated from other gaseous products on a
GC column packed with molecular sieve 5A at a temperature of 30 ºC.
A Faraday cup collector array was used to monitor the masses 2 and 3.
Carbon isotope analysis (ca. 5 mm2 tape sample) was conducted by EAIRMS using a combustion furnace, reduction furnace and GC oven temperature of 1000, 600 and 90 ºC, respectively. Oxygen isotope analysis
(ca. 22 mm2 tape sample) was conducted by total conversion at 1080 ºC
in a quartz reactor tube lined with a glassy carbon film, filled to a height
of 170 mm with glassy carbon chips and topped with a layer (10 mm
deep) of 50% nickelised carbon. Carbon monoxide and nitrogen were
separated on a GC column packed with molecular sieve 5A at a temperature of 50 ºC.
Excellent IRMS results were obtained. For the three different
brands investigated, commercial samples from the same tape product
acquired at different times from one commerical outlet and at one time
from different outlets could be discriminated on the basis of a combination of the δ13C and δ2H results. Alternatively, for the two oxygen
containing brands, samples could easily be discriminated using a combination of the δ13C and δ18O results. This could be done using either
results for the full tape, the glue or for the backing material. Especially
the latter is of great value since this means that, together with the glue,
all other debris (blood, hairs, fibers) can easily be removed.
Discussion: Comparing IRMS with ICPMS an interesting characteristic is that whereas ICPMS focuses mostly on trace elements and is
therefore inherently more sensitive to contamination, IRMS focuses on
the abundant elements (H, C and O) in organic chemical samples and
therefore is less sensitive to contamination problems. The above results
* Presenting Author
demonstrate the strong potential of novel forensic techniques such as
(LA-)ICPMS and IRMS for forensic investigation of packaging tapes. In
our expectation this will prove to be only one example of a wider
forensic application of these techniques for forensic material comparisons.
B168 Capillary Electrophoresis Analysis of a
Wide Variety of Seized Drugs on the
Same Dynamically Coated Capillary
Tape, ICPMS, IRMS
Ira S. Lurie, BA, MS*, Drug Enforcement Administration, Special
Testing and Research Laboratory, 22624 Dulles Summit Court, Dulles,
VA 20166
B167 Elucidating the Relative Dependence of
Propellant Pattern-Based Muzzle-to-Target
Distance Determinations on Variables of
Weapon and Ammunition
After attending this presentation, attendees will have learned a
simple approach for the capillary electrophoresis analysis of a wide
variety of seized drugs.
The methodology presented will greatly simplify the implementation of capillary electrophoresis for a forensic drug laboratory.
Methodology is presented for the capillary electrophoresis analysis
of a wide variety of seized drugs on a single capillary, using multiple run
buffers and diode array UV detection. Drug types analyzed include
phenethylamines, cocaine, heroin, oxycodone, morphine, LSD, psilocybin, opium, and GHB-GBL. Both qualitative and quantitative analyses
are achieved using run buffers than contain additives that provide for
secondary equilibrium and/or dynamic coating of the capillary. Dynamic
coating of the capillary surface is accomplished by rapid flushes of 0.1N
sodium hydroxide/water, buffer containing polycation coating reagent,
and a buffer containing polyanionic coating reagent (with or without
cyclodextrin(s)) or a micellar coating reagent. Dynamic coating with a
polyanionic coating reagent is used for the analysis of moderately basic
seized drugs and moderately basic adulterants. The use of cyclodextrin
in the run buffer not only allows for chiral analysis, but also greatly
enhances separation selectivity for achiral solutes. A dynamically coated
capillary with micellar coating reagent allows for the analysis of neutral
and acid drugs (GHB, GBL and weakly basic, acidic and neutral adulterants). The above approach, which gives rise to a relatively high and
robust electroosmotic flow at pH’s below 7, allows for rapid, precise,
and reproducible separations. For the various drugs examined, excellent
linearity and migration time precision and good peak area precision
(external and internal standard) is obtained. Quantitative results for synthetic mixtures and seized drugs agree with actual values, and also with
results independently determined by other techniques. Both qualitative
and quantitative analyses are greatly enhanced by the use of diode array
UV detection which allows for automation of both library searches and
peak purity.
Jack Hietpas, BS, Peter J. Diaczuk, BS, Jacqueline Speir, BS, and
Peter R. De Forest, DCrim*, John Jay College of Criminal Justice,
445 West 59th Street, New York, NY 10019
The attendee will explore factors that influence the accuracy of
muzzle-to-target distance determinations based on target-deposited propellant particle patterns.
Estimates of the distance from the muzzle of a gun to a nearby
target at the moment of discharge, based on target-deposited propellant
particle patterns (PPP), have figured importantly in shooting investigations for many years. For weapons firing a single projectile, viz., rifles
and pistols as opposed to pellet firing shotguns, these estimates are based
on an analysis of patterns resulting from the deposit of ammunition
derived residues emanating from the muzzle and forcibly deposited on
the target. A major part of this pattern is the PPP. For the purposes of this
study only the PPP is considered. Metallic deposits such as those contributed by shaved and vaporized lead are not considered. This is also
true of deposits due to lubricants and other residues in the barrel. We
hypothesize that these non-PPP contributions are too dependent on the
condition of the barrel and the nature of the ammunition used for prior
shots. The PPP analysis and interpretation require a comparison of the
evidence pattern with a series of test patterns produced on a surface or
substrate similar to the questioned or evidence surface. Ideally, the test
shots are made using the same weapon and ammunition.
In many casework situations no weapon is recovered, although the
ammunition used is known or can be ascertained. Here caution may
dictate that no attempt be made to estimate the muzzle-to-target distance.
However, we postulate that in situations where the ammunition used
during the shooting event can be identified and examples acquired, carefully designed test firings can assist in making reliable estimates.
For this paper it was hypothesized that the most critical variable
affecting the pattern is the ammunition. It was further hypothesized that
nuances in variables related to weapon parameters are relatively less significant. The research in the present paper was designed to test these two
hypotheses. Experimentation involving replicate test firings and the digitization of the resulting PPPs produced the data which, was analyzed.
Following digitization, each propellant particle pattern was characterized. “Image J,” public domain software from the National Institutes
of Health was used to determine the Cartesian coordinates for each propellant particle in a pattern. These data were input into the software
package Math Cad (Mathsoft®) which was used for the analysis and
characterization of the pattern produced by each shot. This enabled the
objective comparison of PPPs and allowed any significant pattern differences resulting from the deliberate altering of gun barrel variables to
be evaluated in relation to stochasic differences among replicate test
shots. The effects of batch-to-batch variations in ammunition were also
evaluated preliminarily. Circumstances where reasonable estimates of
the muzzle-to-target distances can be made in the absence of the actual
weapon used will be described.
Muzzle-to-Target Distance, Propellant Particle Patterns, Gun Shot
Residue
* Presenting Author
Seized Drugs, Capillary Electrophoresis, Dynamically Coated
Capillaries
B169 Defining Appropriate Peak Ratios for
the STR DNA Analysis of Pristine
Single Source Samples
John H. Ryan, PhD*, Myriad Genetic Laboratories, 320 Wakara Way,
Salt Lake City, UT 84108; Dennis Loockerman, PhD, Crime
Laboratory Services, Building B, 5805 North Lamar Boulevard, Austin,
TX 78752; Jeffrey Barrus, BS and Brian Ward, PhD, Myriad Genetic
Laboratories, 320 Wakara Way, Salt Lake City, UT 84108
humanity by defining appropriate guidelines for DNA data analysis of
pristine single source DNA samples.
In casework DNA samples, the relative peak height or area of heterozygous STR alleles are useful indicators to detect the presence of a
potential sample mixture. However, in pristine single source DNA
samples, sample mixtures are not common or expected. To date,
defining acceptable peak ratios in pristine single source DNA samples
has largely been based on the rules used to define mixtures in casework
samples. These peak ratios are often defined as the height of the secondary peak in a heterozygous marker to be greater than or equal to 60%
107
of the height of the main peak. While such a conservative peak ratio
may be appropriate for defining mixtures in casework type DNA
samples it is potentially restrictive for pristine single source DNA
samples. A defined peak ratio of 60% can potentially create unnecessary
work for the DNA data analyst by excluding scientifically robust data.
The acceptable peak ratio for pristine single source DNA samples should
be defined by the accuracy of the resulting STR DNA profile.
We will present the results of a study comparing STR profiles generated from over 130,000 single source DNA samples, in which the
acceptable peak ratio will be changed from 60% to a series of more
liberal peak ratios. The resulting STR profiles generated from each
defined peak ratio will be checked for accuracy. The results of this study
will enable the forensic community to define an appropriate peak ratio
for heterozygous STR markers, of pristine single source DNA samples,
that will produce an accurate profile. In addition, the results of this study
will quantify the amount of work saved by employing a more liberal
peak ratio for heterozygous STR markers of pristine single source DNA
samples.
A mobile phase flow rate of 0.20 ml/min was utilized, and a solvent gradient was used to achieve better separation. Mass spectrometer parameters, which were optimized for sensitivity, included the drying gas
flow rate for the electrospray interfase (ESI) at 12.0 L/min, and the ESI
spray chamber temperature at 350 ºC.
A number of textile fibers were extracted and the dyes identified
using the LC-MS method. Mass spectra of standard dyes and extracted
dyes will be presented and discussed. Dye identification is based on
multi-analyte ions, including various molecule-related ions and
fragment ions. Other advantages of LC-MS, such as isotope information
and the comparison of negative and positive ion spectra, will be discussed.
Peak Imbalance, STR Profiles, Single Source DNA Samples
Guy N. Rutty, MD, MBBS*, and Ali Abbas, Division of Forensic
Pathology, University of Leicester, Leicester Royal Infirmary, Robert
Kilpatrick Building, PO Box 65, Leicester, LE2 7LX, United Kingdom
B170 Fiber Dye Identification by High
Performance Liquid ChromatographyMass Spectrometry (HPLC-MS)
Min Huang, PhD*, Jehuda Yinon, PhD, and Michael E. Sigman, PhD,
National Center for Forensic Science, University of Central Florida,
PO Box 162367, Orlando, FL 32816-2367
The goal of this presentation is to present the progress on the development of a mass spectrometry-based method for the forensic comparison and identification of dyes extracted from textile fibers.
The results of this study offer an improved method for single source
determination in fiber evidence comparisons. Molecular-level information from MS data opens the possibility for dye identification at the
molecular level and the preparation of searchable libraries.
Textile fiber examination is frequently carried out in forensic laboratories to provide physical evidence in crime investigations. An
important part of forensic fiber examination involves the characterization of textile dyestuffs. Compared with currently used methods, such
as ultraviolet, visible, and fluorescence microspectrophotometry,
infrared spectrometry, and high performance liquid chromatography
(HPLC); high performance liquid chromatography-mass spectrometry
(HPLC-MS) provides unique information about the molecular structure
of fiber dyes and makes questioned and known comparisons and/or dye
identification highly discriminating.
Determination of the elution times for extracted dyes is one of the
key issues in applying HPLC-MS to the analysis of dyes. In the absence
of a prior knowledge of a dye’s elution time and/or the major ions
formed from the dye, HPLC-MS analysis may be impractical, if not
impossible, for the analysis of many dyes. This project utilizes tandem
UV-visible absorption and mass spectrometry to facilitate the detection
of dyes separated by HPLC. With visual information about the color of
the textile fibers, the elution of the dyes can be efficiently pinpointed by
a single wavelength UV-visible detector. Diode array detectors offer an
even broader capability for detecting the elution of multiple dyes. In this
way, even dyes that do not show an obvious peak in the total ion current
(TIC) chromatogram can be analyzed. The method also allows extracted
brightening agents and other non-dye components to be distinguished
from the peaks of interest.
An Agilent 1100 MSD quadrupole mass spectrometer equipped
with an electrospray ionization (ESI) source and an Agilent 1100 HPLC
have been used for this study. The instrumentation allows both positive
ions and negative ions to be monitored according to a dye’s tendency to
form negatively or positively charged species. Separation was carried
out with a ZORBAX Eclipse XDB-C18 (2.1 x 150 mm) HPLC column.
108
Textile Fiber, HPLC-MS, Dye Analysis
B171 A Computerized System of Human
Ear Image and Print Identification
The authors intend to present to the forensic scientific community
a computerized system for the identification of the living or dead from
ear images and the identification of offenders from earprints left at
scenes of crime.
This system has 2 principal uses. It can be used to search and
identify an offender from a known database using an earprint left at a
scene of crime. It can also be used to identify an individual purely from
an ear image. This could be used at immigration, customs, CCTV, or
even the search for a wanted individual any where in the world with a
simple photograph been sent to a central database. Therefore the
question as to whether an individual is who they say they are can be
accompanished with this system.
The ear was first used to identify individuals in the late 19th century
when the criminologist Alphonse Bertillion devised a system of identifying individuals using 11 anthropometric measurements of the human
body. These included measuring the height of the individual, as well as
recording the size, shape and position of the ear on the head. However,
the use of ears in human identification was largely abandoned with the
advent of fingerprinting in the early 20th century. In the later part of the
century there was a resurgence of interest in the use of ears to identify
individuals ("earology") although despite the hypothesis that each ear on
each individual is unique, the uptake of this area of identification has
been slow and controversial with many Courts not accepting ear evidence. This problem is confounded by the lack of peer reviewed, scientific publications of methods used for ear identification. To date, only 2
methods have been published for ear images, those of Alexander and
Iannarelli, and there is to our knowledge no peer reviewed published
method for earprint identification which, to date, relies on manual comparison of a print from a scene of crime with a known suspects earprint.
This one-to-one comparison could bias the interpretation.
The Division of Forensic Pathology, University of Leicester,
England, in conjunction with the commercial company K9 Forensic
Service Limited, Northampton, England, undertook a project to produce
a computerised system of ear image and earprint identification. We first
reviewed all available literature on methods used to identify individuals
from their ears and realised a paucity of information and techniques. We
then considered the anatomy, development, racial, genetic and medical
aspects of the ear which may make it unique to an individual. Using volunteers we developed a system for ear image capture using digital photography and collected 800 images (400 right and 400 left ears). All ears
were also printed. This formed the basis of 2 databases (800 images, 800
prints). Random ear images and prints known to be in the databases
where then searched manually using the identified manual methods to
assess ease of use, speed, intra-observer differences and accuracy of
* Presenting Author
identification. All images were then entered into a computer system to
create an ear image and earprint database. The computer system works
by allowing the operator to apply up to 20 points to each image in a
similar way to the fingerprint system. The software then uses the points
to search for the same ear in the database. To enable the programme to
work we had to standardize the placement of the points. This required
the development of a locating grid to be applied to the ear to ensure that
all points, on all ears are placed at the same point. A standardised system
was developed for both ear images and ear prints. In the latter cases this
is, to our knowledge the first such method to be presented and published.
The system which has been developed works for both ear images
and ear prints. It can use complete images or, as often is the case for
prints, partial prints. Although the operator still has to confirm the final
match, the system is considerably more time efficient compared to
manual methods. We have also analysed the differences between left and
right ears, males and females as well as racial differences and produced
a series of standard ideal ear shapes which can be used for facial reconstruction. Finally, by analysis the position of ear piercings we have identified another identifying feature of the ear and shown why earprints are
often incomplete at scenes of crime.
This system has 2 principal uses. It can be used to search and
identify an offender from a known database using an earprint left at a
scene of crime. It can also be used to identify an individual purely from
an ear image. This could be used at immigration, customs, CCTV or
even the search for a wanted individual any where in the world with a
simple photograph been sent to a central database. Therefore the
question as to whether an individual is who they say they are can be
accompanished with this system.
Ear Image, Earprint, Computer
B172 Extractionless GC/MS Analysis of
γ-Hydroxybutyrate and γ-Butyrolactone
With Trifluoroacetic Anhydride and
Heptafluoro-1-butanol From Aqueous
Samples
Alberto J. Sabucedo, PhD*, and Kenneth G. Furton, PhD, Florida
International University, University Park-Department of Chemistry
CP-178, Miami, FL 33199
This presentation will describe the development of a technique
designed for qualitative and quantitative determination of
γ-Hydroxybutyrate (GHB) and indirectly γ-Butyrolactone (GBL) from
aqueous matrices without the need for an organic extraction. GHB is
derivatized in the presence of water without the need for an organic
extraction. This technique offers several advantages such as rapid determination, minimum sample handling, low sample volume, high sensitivity, improved mass spectra for confirmation and indirect determination of GBL concentration.
This may become a general technique in both the toxicology arena
and the drug testing community. The technique offers minimal sample
handling and excellent detection of a commonly abused drug in our
society.
γ-Hydroxybutyrate (GHB) is a DEA Scheduled I drug of abuse
commonly spiked into beverages to incapacitate victims of sexual
assault. GHB is a challenging drug for analysis by GC/MS because of its
small size, charged nature, low volatility and intramolecular esterification leading to γ-butyrolactone (GBL). In this work an extractionless
technique has been developed that allows for the use of an aqueous
sample for direct derivatization. The technique uses a solution of trifluoroacetic anhydride (TFAA) and 2,2,3,3,4,4,4-heptafluoro-1-butanol
(HFB) to derivatize the active hydrogens of GHB. The conversion of
* Presenting Author
GBL to GHB can be forced under alkaline conditions by diluting the
sample in 10 mM borate buffer, pH 12.0. Legally GBL found in beverages intended for human consumption is considered a scheduled substance under current analogue law. Spikes of the two compounds into
several beverage matrices gave quantitative recovery of GHB by
GC/MS. The derivatization produces higher molecular mass products
whose fragmentation pattern provides multiple peaks for confirmation
and quantitation. The concentration of GBL can also be indirectly determined by the method developed by analyzing an aliquot of the same
sample under hydrolytic conditions and in water. Therefore, this extractionless technique is rapid, sensitive and selective for the confirmation
of the presence GHB and GBL in commercial beverages.
The current study shows the utility of an extractionless technique
for the determination of GHB. The technique can be as simple as taking
an aqueous neat sample and performing the derivatization. The dilution
with borate buffer is only necessary if the conversion of any GBL to
GHB is desired by hydrolysis of the lactone. The derivatization was
monitored by full-scan mode to choose the ions with best sensitivity and
selectivity. This derivatization technique produces sufficient high abundance fragments to make the identification by selected ion monitoring
mode (SIM) easier than derivatizations that produce few and low abundance ions such as BSTFA/1%TMCS derivatives of GHB. Both the
esterification and trifluoroacetylation are products of this reaction.
The standard curve has a linear range of 0 to 100 ppm with a correlation coefficient, r = 0.999. A ratio of standard (GHB) to internal
standard (GHB-d6) response was used to correct for variations in the
derivatization and chromatographic processes. A wide range 0 to 1000
ppm standard curve gave a coefficient of correlation of r = 0.996. This
analysis has a limit of detection (S/N=3) of 50 ppb and a limit of quantitation (S/N=10) of 150 ppb. The precision of the assay is excellent
with percent coefficient of variation (%CV) below 5%.
Spike and recovery studies were performed with some common
neat beverage matrices with and without alcohol. The recoveries are
within 11% of the target value. This is impressive since the sample
matrix was not diluted. In addition, the neat rum spiked sample contains
approximately 40% ethanol. The next set of studies involved spiking
both GHB and GBL into the different matrices and comparing their
recoveries against the GHB standard curve. In this set of experiments the
1:10 dilution in 10 mM borate buffer, pH 12 was used to force the
hydrolysis of GBL to GHB. GHB recoveries were on average about
100% and the GBL+GHB spiked matrices were on average 106%. The
use of selected ion monitoring (SIM) mode for the quantitative analysis
increases sensitivity over full-scan mode. The high concentrations
encountered in typical beverages can easily allow the analysis to be performed in full-scan mode. The estimation of GBL concentration present
in an aqueous solution can also be indirectly determined by submitting
an aliquot of the sample to the alkaline hydrolysis and another aliquot to
a simple dilution in water. The difference between the two values is negligible if the substance present is only GHB. If GBL is present than the
two values will differ accordingly. Several urine matrix spikes were performed and the recoveries were within 15% of the target spiked values.
Therefore, this assay may also be used for the determination of GHB in
biological matrices.
Analysis of compounds with similar chemical structures included:
4-aminobutyric acid (GABA), diethylene glycol, 1,4-butanediol and
gamma-butyrolactone (GBL). These were derivatized and monitored to
account for any contribution to the GHB response. GBL was tested
without the hydrolysis conversion step of the method to check the
percent that converts to GHB in the process. The materials were spiked
to a maximum of ten times the upper GHB standard curve concentration.
The spiked concentration of the test compound was read from the GHB
standard curve and the percent response was calculated from the fitted
regression slope of the compound across the range tested. The only compound that gave a significant GHB peak was GBL. The GBL response
was approximately 10% under these conditions across a wide range.
109
Some of the benefits of this method are the lack of an extraction
step, very small sample size, the rich mass spectra obtained upon derivatization providing many abundant ions available to monitor by SIM
mode, increased sensitivity and ability to detect both GHB and GBL in
a complex beverage matrix. The typical analysis involves a significant
dilution to get the concentration in the range of the assay. This dilution
can be performed with either deionized water or the hydrolyzing borate
buffer. The detection of either GHB or GBL in beverages is important in
the forensic community because of the legal ramifications and the
intended illegal use of GBL as an analogue of GHB. In vivo GBL is converted to GHB thus making the presence in beverages an illegal controlled substance similar to GHB. The conversion step of GBL to GHB
allows for chemist not to miss the current trend of spiking GBL in beverages intended for human consumption. The technique requires
minimal sample handling and provides reliable quantitation of GHB
and GBL.
GHB, GBL, Extractionless
110
* Presenting Author
j
DALLAS
C1
Engineering Sciences
Mold Investigations — What is
the Science Behind the Work?
Gary R. Brown, PE*, RT Environmental Services, 215 West Church
Road, King of Prussia, PA 19406
After attending this presentation, attendees will have an understanding of how semi-quantitative techniques are used to assess
microbial growth problems.
Health effects from mold have received increased public and
medical attention. This presentation will impact the forensic community
and/or humanity by increasing the understanding of the limits of what
can and cannot be measured, which is critical to understanding how this
practice area operates.
This paper has several objectives:
• Understanding how microbial growth is measured.
• Understanding how data is interpreted to determine if
amplification is occurring.
• Conveying to the participant how microbial consultants interpret
all the information from a site to reach decisions on microbial
remediation and/or occupancy issues.
The practice of microbial consulting is different from all other environmental practice areas. While the actual mycotoxins associated with
molds are not usually measurable, there are direct associations between
certain types of mold and health symptoms in humans. Gaining an
understanding of this emerging science is fundamental to understanding
which molds are and are not of concern.
Mold, Microbial, Amplification
C2
The Analysis of Darkening
Agents in Indoor Environments
James R. Millette, PhD* and Richard S. Brown, MS, MVA, Inc.,
5500 Oakbrook Parkway, Suite 200, Norcross, GA 30093
information about the microscopical analysis of the causes of darkening
and discoloration in indoor environments.
humanity by demonstrating forensic microscopical studies which show
the dramatic differences between mold, soot and other contaminants that
cause darkening and discoloration in indoor environments, thereby
helping to provide the scientific information necessary for judicial
decisions.
In recent years, toxic mold contamination has spawned a growing
number of complaints and lawsuits from Delaware to California. A
landmark June 1, 2003, Travis County, Texas, jury ruling decided that an
insurance group must pay more than $32 million to a Dripping Springs
family because of sickening black mold damage. In New York, class certification was denied in an $8 billion mold case. Concern over mold in
indoor environments has clearly become a legal/environmental problem,
but the dark discoloration that occurs in residences and workplaces is not
always mold. As illustrated by an MVA project, “A Spot Called Ralph,”
environmental forensic microscopy can provide the scientific data
needed to determine the cause of darkening and discoloration. As
described in South Carolina’s leading newspaper, “A mysterious stain
has appeared in the new carpet at the Lexington County’s Courthouse.
Employees at the courthouse have even given the spot a name ‘Ralph.’“ Not only did the stain appear at the courthouse, but after
cleaning reappeared and grew in size from about a quarter to several feet
* Presenting Author
j
2004
in diameter. Testing of the carpet for mold was negative. Because of the
unknown nature of the stain, the courthouse offices with the spot were
closed as a precaution. A sample of the carpet was delivered to the MVA
laboratory for testing. Analysis by polarized light microscopy (PLM)
and scanning electron microscopy (SEM) showed that the particles
present on the carpet were typical dirt found in offices. A sticky substance holding the particles to the carpet fibers was found to be consistent with corn syrup by Fourier transform infrared (FTIR) spectroscopy. High fructose corn syrup is an ingredient in many soft drinks.
The spot called Ralph was probably a combination of typical office dirt
and a spilled drink that when not completely cleaned simply spread the
sticky substance so the spot appeared to grow.
Black, low temperature combustion soot also causes darkening and
discoloration of indoor environments. The difference between mold and
soot can be easily seen by microscopy. Some information about the
sources of soots can also be determined. Morphology and elemental
analysis of soot aggregates by analytical transmission electron
microscopy (ATEM) using ASTM D6602 - Standard Practice for
Sampling and Testing of Possible Carbon Black Fugitive Emissions or
Other Environmental Particulate, or Both, (American Society for Testing
and Materials, 2000) can be used to compare with soots from known
sources, such as carbon black, fireplace, candles, oil and gas furnaces.
Mold, Soot, Microscopy
C3
The Toxicology of Molds
James S. Smith, Jr., PhD*, Oak Creek, Inc., 60 Oak Creek, Buxton, ME
04093-6616
After attending this presentation, attendees will have a better understanding of: 1) mold growth and the development of mycotoxins; 2)
mold-related diseases in humans and other animals; 3) detection of mold
and mycotoxin exposure; and 4) the assessment of risk posed by molds
and mycotoxins. Dr. Smith will use a case study to illustrate the difficulties encountered in defining mold-related injury.
Over the last few years, insurance companies, landlords, tenants,
home owners and others have become acutely aware of the potential
health effects and financial liability posed by molds in indoor air. Often
such health effects and liability are assumed regardless of the specific
mold or molds encountered. This presentation will impact the forensic
community and/or humanity by describing the toxicology of specific
molds, which is a necessary prerequisite to the evaluation of their
potential to adversely impact indoor air quality and pose a human health
risk worthy of study and remediation. This presentation provides basic
toxicological information about molds required for the evaluation of
indoor air environments and understanding the potential for adverse
effects in people.
This paper has three objectives: (1) to provide an overview of the
toxicology of molds; (2) to identify conditions leading to mold and
mycotoxin growth, development, and exposure; (3) to describe adverse
health effects attributed to mold and mycotoxin exposure; and (4) to
describe the difficulties inherent in demonstrating a mold-related injury.
Recent multi-million dollar jury awards in high profile legal cases
involving mold-related injury captured the attention of insurance companies, regulators, environmental firms, realtors, landlords, and the individual homeowner. In the midst of an explosion in the growth of moldrelated assessment and remediation services, there is too little emphasis
placed on recognizing mold related injury.
Molds are fungi, ubiquitous organisms that make up approximately
25% of earth’s living matter. They play an important role in the
breakdown of organic matter like leaves, wood, and plant debris. To
111
flourish, molds require moderate temperatures (i.e., between 40 and
100°F), a nutrient base (such as the cellulose in wood or paper), and
moisture. People become exposed to mold and/or mold-toxins (i.e.,
mycotoxins) through the consumption of mold and/or mycotoxin in
foods, having skin contact with mold and/or mycotoxin, and through
inhalation of mold spores/fragments and/or mycotoxins. Exposure to
mold and/or mycotoxins is associated with immunosuppression, immunodepression, emesis and diarrhea, weight loss, nervous disorders, cardiovascular alterations, skin toxicity, decreased reproductive capacity,
bone marrow damage, flu-like symptoms, including sore throat and
headache, dizziness, dermatitis, fatigue, and general malaise.
Asthmatics tend to be particularly sensitive to mold exposure, with 10 to
32% of all asthmatics showing some sensitivity. Perhaps the most sensitive group of people is those with compromised immune systems.
It is often useful to think of the adverse health effects associated
with molds as falling into two basic categories; those caused by exposure
to the mold organism itself and those caused by exposure to a mold-produced toxin or mycotoxin. In the former category, direct exposure to
molds is known to illicit a wide variety of responses from humans and
other animals. These responses include diseases characterized by the
growth of mold on tissues (e.g.; athlete’s foot) and secondary health
effects (e.g.; increased incidence of disease), which may be related to the
suppression or depression of immune system function. In the latter category, doctors have used certain mycotoxins to treat bacterial infections
(e.g.; penicillin), while inadvertent exposure to other mycotoxins is
clearly associated with the development of cancer and even death.
Recognizing mold-related injuries is critical to the identification of
mold problems. This is because nearly half of the buildings with moldrelated indoor air quality problems do not present visible signs of mold
growth. Consequently, environmental companies often recommend
thorough and competent testing to evaluate indoor environments for
mold. Air sampling alone, however, will not provide sufficient evidence
to indicate a mold problem and because there are no official standards or
guidelines for regulating molds in indoor air, determining that a mold
problem exists can be problematic even with good indoor air sampling
information. Consequently, the most important feature of an assessment
of a mold problem is the evaluation of mold-related injuries. In this presentation, Dr. Smith will present an overview of mold toxicology with
special focus on the difficulties inherent in demonstrating mold-related
injury.
mental investigations and governmental regulations. In this example,
the forensic environmental microscopist was asked to determine the size
of fine particles available in the lead ore concentrate that could potentially become airborne and be inhaled or ingested. Items of clothing,
including a respirator worn during a typical workday, were examined for
the presence of fine (less than 2.5 micrometer) particles containing lead.
The handling of the lead ore concentrate was simulated in a controlled,
closed environment to observe and collect samples of any dust produced.
The experiment was videotaped and air samples were collected to
determine what the airborne particle size distribution could be.
Sections from the submitted clothing and the respirator filter were
placed in a solution of water and a dispersant. Particles were separated
from their respective substrates by ultrasonicating the sample sections in
the water/dispersant solution for several minutes. The resulting particle
suspension was deposited onto a polished carbon substrate for analysis
by SEM-EDS. Examples of lead-rich particulate were documented. No
quantitative measure of the particulate released from the clothing and the
respirator was performed.
A glove box was prepared to simulate the handling of the lead ore
concentrate. Lead ore concentrate was transferred from one container to
another by scooping, pouring and sweeping within the glove box. The
glove box was equipped with a clear viewing area so that the handling
of the lead ore concentrate could be video taped using a focused beam of
light. Particles that entered the focused beam of light were highlighted
in the same way as ambient dust is highlighted by a ray of light coming
through a crack in an otherwise darkened room. This effect where the
fine suspended dust particles are highlighted is known as the Tyndall
effect; the reflection of light by particles suspended in a transparent
medium. Suspended particles were collected with air filter cassettes
present in the glove box during the simulation. Particle sizing over the
range of 0.5 to 10 micrometers was performed utilizing automated
scanning electron microscopy on the collected air filter samples to
determine the size distribution of the lead ore dust generated during the
simulation.
Forensic Environmental Microscopy, Lead Ore, SEM
C5
NIST Standard Reference Materials
(SRMs) for Forensic Measurements
and Analysis
Mold, Toxicology, Toxic Molds
C4
Automated Scanning Electron
Microscopy of Lead Ore Dust
Richard S. Brown, MS*, MVA, Inc., 5500 Oakbrook Parkway,
Suite 200, Norcross, GA 30093
The goal of this presentation is to present to the environmental
forensic community the use of scanning electron microscopy (SEMEDS) as a tool to investigate the particle size distribution of lead ore dust
(galena) in a controlled, closed experiment and to determine the
presence of lead particles on submitted items of clothing.
humanity by demonstrating the techniques used for criminal forensic
science which are applicable to environmental law and litigation.
Attendees will gain an understanding of how particle sizing is performed
utilizing an automated scanning electron microscope after dust collection from airborne particulates. Sampling procedures for examining
clothing and a respirator filter will be shown for the detection of a nonwater soluble, high atomic number particulate such as lead sulfide. The
size distribution supported claims by the plaintiffs that they had been
exposed to respirable lead dust during their workday.
Environmental forensic microscopy is the application of
microscopy to the collection, analysis and identification of small
particles and the interpretation of any results as they pertain to environ112
Mario J. Cellarosi, BA, MS*, National Institute of Standards and
Technology, Gaithersburg, MD 20899-2320
After attending this presentation, attendees will learn how to use
NIST Web-based resources and available NIST Certified Standard
Reference Materials which provide the benchmarks of precision,
accuracy, and traceability to validate measurements to be used as evidence.
humanity by describing to the forensic community the valuable
resources available from the National Institute of Standards and
Technology (NIST) on Standard Reference Materials which can be used
to validate measurements in forensic evidence.
Presently available NIST Certified Standard Reference Materials
(SRMs) related to measurements and characterization of specimens for
linkage to forensic evidence are described. The NIST SRMs Web
presence and its navigation techniques are also demonstrated.
This paper discusses physical and chemical properties of certified
NIST SRMs related to measurements in the identification and/or comparison of specimens to be linked to forensic evidence. NIST supports
accurate and compatible measurements by providing over 1300 Certified
SRMs with well-characterized composition and/or properties. These
SRMs are used to perform instrument calibrations in situ as part of
overall quality assurance programs, to verify the accuracy of specific
measurements, and to support the development of new measurement
* Presenting Author
methods. NIST SRMs are currently available for use in areas such as
industrial materials production and analysis, environmental analysis,
food and agriculture, radioactivity, health measurements and basic measurements in science and metrology. Each SRM is supplied with a
Certificate of Analysis. Along with other standardization organizations
methods and procedures, such as ASTM and ANSI, NIST has published
many articles and practice guides that describe the development,
analysis and use of SRMs. These SRMs provide the benchmarks of precision, accuracy, and traceability which validate the evidence.
The measurement of physical, optical and chemical properties of
materials samples are often employed to identify the type of material
and/or application. Measurements of material properties can be used to
track and identify the original producer, the date or period of manufacture and the intended use or application for the material or product.
For example, property or chemical measurements and/or the evaluation
of materials or product characteristics, in addition to visual markings if
present, can establish a link in the chain from producer, fabricator, distributor, vendor, end-use or application, down to a specific geographical
area or sample origin.
In the measurement of properties, chemical composition, or characteristics of a material, accuracy and uncertainty terms and traceability
statements are of paramount importance in the validation of evidence for
forensic investigations. These concepts must be used correctly to avoid
possible confusion and inadmissibility of evidence. SRMs and the associated Certificate or Certificate of Analysis documentation state the
intended purpose and application of a particular SRM, its certified
property value(s) with associated uncertainty (ies), and present technical
information deemed necessary for its proper use. The uncertainty
attached to a certified value is especially important as it represents a
quantity which characterizes the range of values within which the true
value is asserted to lie with a stated level of confidence. A NIST SRM
certificate bears the logo of the U.S. Department of Commerce, the name
of NIST as certifying body, and the name and title of the NIST officer
authorized to accept responsibility for its contents. In addition to the certified values, the SRM certificate may contain references and/or other
pertinent information and data. Certified values for SRMs with their
associated uncertainties, in applicable situations, insure the integrity and
the validation of forensic measurements. NIST-certified values are
obtained by one or more of the following measurement modes: 1) A
definitive (or primary) method using specialized instrumentation
capable of high accuracy and precision and whose errors have been thoroughly investigated and corrected; 2) Two or more independent
methods at NIST using commercial instrumentation that is calibration
based and with differing sources of systematic errors; or, 3)
Interlaboratory data from selected laboratories using multiple methods
and SRMs as controls. However, the sources of error with the latter
mode will generally result in uncertainties greater than those for the
other two modes.
There are a number of measurement methodologies related to the
determination of materials properties and/or chemical composition. For
instance, chemical composition methods cover basic “wet chemistry”
procedures and other very sophisticated techniques, which utilize atomic
and radiation physics principles, and nuclear interactions that require
complex and expensive apparatus. Fortunately, a number of SRMs
having components comparable with those of the materials to be evaluated have been established. These SRMs and associated methods or
standard procedures are available for equipment calibrations.
This paper will discuss and illustrate the use of a number of SRMs
of interest to the forensic community. The discussion will encompass
measurement practices, methods, standards, and precision and accuracy
considerations to be taken into account for the measurement methodologies employed. This paper will also provide insights on the future
needs for SRMs to be utilized in measurements and materials characterization.
Standards, Reference Materials, Calibrations
* Presenting Author
C6
Recent Advances in High Resolution
FT-ICR Mass Spectrometry and Their
Applications to Environmental Forensics
Ryan P. Rodgers, PhD*, Geoffrey C. Klein, BS, and Lateefah A.
Stanford, BS, Department of Chemistry and Biochemistry, Florida State
University, 1800 East Paul Dirac Drive, Tallahassee, FL 32310;
Christopher L. Hendrickson, PhD, National High Magnetic Field Lab,
Florida State University, 1800 East Paul Dirac Drive, Tallahassee, FL
32310; Alan G. Marshall, PhD, Department of Chemistry and
Biochemistry, Florida State University, 1800 East Paul Dirac Drive,
Tallahassee, FL 32310
After attending this presentation, attendees will understand the fundamentals of high resolution FT-ICR mass spectrometry. Fundamental
limitations of traditional analytical techniques, and how FT-ICR MS
overcomes many of those limitations will be discussed.
humanity by describing how the amount of information that can be
obtained from complex environmental samples has drastically increased
as a result of FT-ICR MS applications to environmental forensics.
Thousands of different compounds may now be identified and used for
fingerprint-based identification of heavy crude oil or other complex contaminants.
Traditional analytical techniques such as Liquid Chromatography
(LC) and Gas Chromatography (GC) have inadequate chromatographic
resolution for the baseline separation of all species present in complex
mixtures composed of hundreds or even thousands of individual components. As a result, combined techniques (such as GC-MS and LC-MS) are
ineffective due to co-eluting species that complicate the mass spectrum
and hinder mass spectral interpretation and component identification
through traditional library based searches. This is especially true for
complex materials that have undergone abiotic or biotic modification in
the environment. Furthermore, mass spectrometers commonly employed
for GC-MS and LC-MS are low resolution/low mass accuracy quadrupole
mass filters or quadrupole ion trap type mass analyzers that are unable to
adequately resolve complex mixtures for individual component identification. Even if the traditional analytical techniques succeed in both separating and resolving an individual species in a complex mixture, the low
mass accuracy provided by the commonly employed mass spectrometers
prevents elemental composition assignment. Fourier Transform Ion
Cyclotron Resonance Mass Spectrometry (FT-ICR MS) benefits from
ultra-high mass resolving power (greater than one million), high mass
accuracy (less than 1 ppm), and rapid analysis; these attributes make it an
attractive alternative for the analysis of complex mixtures such as crude
oil. For example, we recently resolved almost 20,000 different elemental
compositions in a single positive-ion electrospray FT-ICR mass spectrum
of a heavy crude oil.
Here we report environmental forensic applications of FT-ICR MS
in the compositional analysis of heavy crudes, their water-soluble
fraction, and abiotic changes in the crude oil polar species as a function
of increased weathering in both soil and water (simulated sea water).
High resolution (>300,000) and high mass accuracy (sub-ppm) mass
spectra of electrospray (ESI) ionized samples are obtained on a homebuilt 9.4T ESI FT-ICR MS equipped with a MIDAS data station. Crude
oil standards consisted of thousands of peaks over hundreds of compound classes. Five geochemically different crudes were analyzed in an
effort to differentiate them from one another based on their compositions
provided from the FT-ICR MS analysis. The crudes showed significant
differences that directly relate to their geochemical source histories. We
also report the compositional determination of water-soluble organics
leached from a heavy crude oil. Contrary to previous reports in the literature, the composition of the water-soluble species is very complex
and is composed of both the expected CO2 series (carboxylic acids) as
well as higher oxygen containing hydrocarbons and many SOX species.
113
Similar weathering experiments in sterilized soil will also be presented.
Relevant instrumental advances will be discussed that both expand the
compound classes that can be identified and make the analysis more
informative and routine. Advances discussed include selective ion accumulation, a new octapole ion accumulator that increases the efficiency of
ion ejection, and a new field desorption/ionization/atmospheric pressure
ion source. Work supported by ExxonMobil Research and Engineering,
NJ, NSF (CHE-99-09502), FSU, and the National High Magnetic Field
Laboratory in Tallahassee, FL.
High Resolution Mass Spectrometry, Environmental Forensics,
FT-ICR MS
C7
To Daubert or Not to Daubert?
That Has Become THE Question
James S. Smith, PhD*, Trillium, Inc., 28 Grace’s Drive, Coatesville,
PA 19320
After attending this presentation, attendees will understand the use
or misuse of a Daubert hearing.
humanity by warning expert witnesses and their attorneys about the use
of a Daubert hearing and the gatekeeping, or lack thereof, during trial.
The cost, quite often, is a loss.
The Daubert v. Merrill Dow Pharmaceuticals, Inc. Supreme Court
decision is 10 years old. It established guidelines for the admissibility
of scientific evidence in Federal courts and it has trickled down to most
state courts. The judge must be gatekeeper and determine what is
reliable evidence and what is junk science. General Electric v. Joiner in
1997 and Kumho Tire v. Carmichael in 1999 were Supreme Court cases
expanding the judge’s gatekeeper role over all expert testimony. In the
Kumho Tire decision, the Supreme Court specifically pointed out that
opinion is not science just because an “expert” says so. In other words,
when an expert witness starts a sentence with “In my experience....,”
then the gatekeeper should slam the gate shut. Simply stated, the
Daubert decision requires that opinions of expert witnesses be examined
with the same scientific rigor as scientific statements made outside the
courtroom.
However, Daubert hearings occur only when either the plaintiff or
the defendant makes a request for the hearing. The following case illustrates the use of the Daubert hearing, and how the judge was not a gatekeeper.
The plaintiff’s expert has good credentials to be involved in an
environmental litigation and produces a lengthy, detailed expert report
following the Federal Rules of Evidence. The defendant’s expert does
not have good scientific credentials and produces a two page letter in
response to the plaintiff’s expert report. The letter is full of blatant inaccuracies about the known published science concerning the contaminants of interest in this litigation.
Depositions of these two experts are taken. The plaintiff’s expert
can and does support the expert report with literature references and documentation. The defendant’s expert either hasn’t studied the various
issues or cannot make a definitive statement. However, this expert is
positive that the plaintiff’s expert is wrong.
Now, the surprise is that the defense wants to exclude the plaintiff’s
expert based on the unreliability of the scientific approach used in this
case. This is bolstered by a decision by another judge to disallow a
portion of an affidavit written in another case in another state. What followed was a rigorous questioning of the expert fro the plaintiff, by the
defense attorney, the judge, and the plaintiff’s attorney. If the judge
decides to close the gate then this expert’s testifying days are over. The
expert does survive, but there is no doubt about what that expert will
present at trial because it has been cast in stainless steel via the expert
report, deposition and Daubert hearing.
114
What about the weak, so-called expert for the defense? At trial the
plaintiff’s attorney will kill him over his inaccurate letter report and lack
of scientific knowledge. But, at trial, the testimony has nothing to do
with the letter report or deposition question and answers. A surprise
demonstration is aimed at the weakest kink in the plaintiff’s case, which
was really not that weak. Objections do not carry the day and there is
nothing available in the trial testimony to allow for a rigorous cross
examination.
The Daubert challenge by one side should automatically force a
challenge of both experts. Rigor should not be applied to one side
without being applied to the other. Scientific evidence was not fairly
presented nor was the gatekeeper aware of the (mis)use of a Daubert
hearing to tip the scales of justice.
Daubert, Kumho Tire, Gatekeeper
C8
A Study of Several Case Histories of
Successful Method Detection Limit
(MDL) Studies Consistent With
40CFR136, Appendix B
James E. Norris, MS*, Severn Trent Laboratories, 900 Lakeside Drive,
Mobile, AL 36693
After attending this presentation, attendees will have a knowledge
of the utility of the Method Detection Limit determination under
40CFR136, Appendix B.
humanity by demonstrating a deeper understanding of the limits of
USEPA methods in analyses of various substrates.
This presentation addresses several of the 15 Method Detection
Limit studies conducted by the author for industrial facilities in four different states. All were successful from both a technical and a regulatory
perspective.
The strategic objective of each study was straightforward: to
determine a matrix-specific, analyte-specific, method detection limit
(MDL) in an industrial wastewater discharge and to accomplish this in a
manner consistent with USEPA’s methodology given at 40CFR136,
Apdx B. Prior to each study, the responsible State (or Federal) permitting agency had been informed of the need and the intent to execute
such a study and in no case did the responsible agency veto the necessity
of conducting such a study.
This presentation will address the studies of several analytes and
methodologies (organic compounds, GC/ECD and GC/MS), non-metals
(e.g., Total Cyanide distillation with automated colorimetry), metals (by
ICP/GFAA), and pesticides (GC/ECD, GC/MS). A comparison will be
made to demonstrate the often dramatic differences in EPA-published
MDLs versus those defined in real-world industrial wastewater matrices.
One implication of the matrix-specific MDL is demonstrated by a
significant drop in the number of NPDES permit limitation violations.
The introduction by USEPA of the concept of the Minimum Level
(ML) as, effectively, a limit of quantitation (LOQ) will be shown to
further ease the compliance burden on permitted industries. Unhappily,
the EPA has not incorporated the ML concept into binding regulation (as
is the case with the MDL) but has presented the ML as mere guidance.
Some states (e.g., Texas) have adopted the ML concept and incorporated
it into their state water-permitting regulations.
Interestingly, all new and updated 40CFR136 methods include not
only the MDL but also the ML for the analytical method. In this paper,
the most frequently evaluated MDLs have been one form or other of
cyanide (e.g., Total Cyanide, Cyanide-Amenable-to-Chlorination, Weak
Acid Dissociable Cyanide).
Impetus to conduct such MDL (and ML) studies resides in the
growing tendency of both USEPA and State agencies to set discharge
limits below the limits of detection. The success of the various LEAF
* Presenting Author
(Legal Environmental Advocacy Fund) lawsuits and the rapidly
expanding program of re-classification of streams and rivers (leading to
Total Maximum Daily Loads) have resulted in water quality driven mass
discharge limits which translate into immeasurably low concentrations
in wastewater discharges.
Unhappily, many NPDES permit writers do not seem to understand
that compliance with a mass-based limit is contingent not only on flow
(thus, total mass of wastewater) but upon a measurement of concentration of the offending species. This aspect of the compliance problem
will also be addressed in this presentation.
Analytical, Detection Limits, Matrix-Specific
C9
Engineered Containment Systems for
Radioactive and Hazardous Wastes —
Do Current Approaches Present
Opportunities for Future
Environmental Forensics?
James H. Clarke, PhD*, Vanderbilt University, Department of Civil
and Environmental Engineering, PO Box 1831, Station B, Nashville,
TN 37235; Ann N. Clarke, PhD, ANC Associates, Inc., 7101
Executive Center Drive, Brentwood, TN 37027; Lorne G. Everett, PhD,
Shaw Stone & Webster, 3700 State Street, Suite 350, Santa Barbara,
CA 93105
The goal of this presentation is to familiarize the audience with the
current approach to long-term containment of wastes, the failure of
which may require future forensic investigation.
humanity by demonstrating the limitations of current containment
systems, some of which may result in the need for future forensic investigations.
The field of environmental forensics has evolved largely as a result
of past chemical and waste management practices that have resulted in
large scale contamination of surface and subsurface environmental
media (surface water and groundwater, soils and sediments). When it
became known that these practices were affecting valuable environmental resources, steps were initiated to both restore the contaminated
media to an appropriate degree, based on risk and future land use, and to
adopt improved standards and practices for the ongoing management of
wastes and cleanup residuals. As a result of major technical and economic limitations on implementing remediation, a large volume of these
contaminated media will be contained in engineered systems either in
situ, using surface and possibly subsurface barriers, or excavated and
placed into new engineered containment systems (disposal cells). Given
the lifetimes of the radioactive and hazardous constituents (hundreds or
thousands of years to essentially forever for stable toxic metals), these
engineered systems will have to be monitored and maintained, with
ongoing access restrictions (institutional controls), for very long time
horizons.
The primary function of an engineered containment system is to
prevent radioactive and/or hazardous constituents from migrating to
potential exposure points. Institutional controls are also needed insure
ongoing monitoring and maintenance and to prevent direct contact with
the contaminated media. The design approaches that have evolved rely
on a primary barrier system whose main function is to keep infiltrating
rainwater and possibly groundwater from contacting the contaminated
materials and wastes that are being isolated and transporting constituents
to the environment. This primary barrier, in a surface barrier system for
example, typically consists of a compacted soil layer, sometimes augmented with bentonite, together with a synthetic membrane (geo-membrane) to keep the soil from desiccating and cracking and to provide an
additional layer of protection. Over the years, as experience has been
gained, additional layers have been added to protect the primary barrier
* Presenting Author
system from the adverse impact of natural processes such as erosion and
bio-intrusion that can compromise the performance of the primary
barrier system. The current design approach typically contains several
layers and is very expensive to construct (typically several hundred
thousand dollars an acre). In some cases, where gases are of concern
(e.g., methane, radon), gas barriers or collection systems are employed
as well.
While our experience with the current state of the art approaches to
system design is limited to a few decades at best, observations of system
performance and data are beginning to emerge. These data and observations suggest that there is merit to re-evaluating our current approach. In
particular, design approaches that can accommodate a certain degree of
environmental change and that do not have to rely on resisting natural
processes are generating a great deal of interest.
Also, progress has been hampered, to varying degrees, by the
current regulations that call for prescriptive designs (that may not be the
best in certain environments), i.e., requiring monitoring of the saturated
zone and only thirty (30) years of post closure monitoring and maintenance for facilities regulated under the Resource Conservation and
Recovery Act (RCRA). In many cases, particularly at the Department of
Energy sites, extensive vadose zones are present that provide separations
of hundreds of feet between the engineered containment system and the
top of the saturated zone. Monitoring of both the vadose zone and the
system itself could provide valuable early warning information concerning system performance.
The authors have spent many years working on both Superfund
sites and the investigation and restoration of former nuclear weapons
production facilities and are currently evaluating system design and
monitoring approaches at a number of sites. The time appears appropriate to determine: the data needed to evaluate the performance of the
sites over time; what tools are needed to collect the data; and what needs
to be built into the design to help ensure the data are available.
Examples will also be provided that illustrate the shortcomings of the
current approach and alternative design and monitoring approaches that
could provide improved and more cost effective protection over long
time horizons.
Long Term Containment, Monitoring Needs, Design Needs
C10
Western Precious Metal Ingots —
The Good, the Bad, and the Ugly
Fred N. Holabird, BA*, Holabird Mining, Environmental & Historical
Consultants, 3555 Airway Drive #308, Reno, NV 89511; David C.
Fitch, MS, PO Box 70547, Reno, NV 89570; Robert D. Evans, BS,
1290 Flint Ridge Road, Hopewell, OH 43746
scope of knowledge necessary in the identification of fake precious
metal ingots.
humanity by demonstrating breakthrough technology for determining
real versus fake historical or modern precious metal ingots.
Historical precious metal ingots have changed hands for millions of
dollars between bankers, institutions and collectors. Elaborate fakes
have entered the market place, causing significant unrest. Experts have
generally been unable to identify fakes until now. Using the gold ingots
from the 1857 wreck of the SS Central America as a baseline, studies for
the past five years have generated new methodology for determining real
historical ingots from fakes.
Scanning Electron Microscopy with Energy Dispersant X-Ray
Spectrometry backscatter scans show original textural features of the
metallurgical product that can be compared to modern metallurgical
pours. Laser Ablation-Inductively Coupled Plasma Mass Spectrometry
analyzing major and trace element chemistry is used to compare original
historical ingots with questioned pieces. A thorough understanding of the
115
historical record associated with the companies that poured these ingots
is necessary to further interpret the data.
Using this new methodology, several ingots have been determined
to be fake, including one important ingot in the Smithsonian Collection.
Precious Metal, Gold and Silver, Fraud
C11
Biomechanical Determinants of
Injuries From Low-Level Falls
John R. Brault, MS*, John C. Gardiner, PhD, Dennis D. Chimich,
MSc, PEng, and Gunter P. Siegmund, PhD, MacInnis Engineering
Associates, Inc., 21076 Bake Parkway, Suite 106, Lake Forest,
CA 92630
The goal of this presentation is to present a review of published scientific research in order to enhance the ability of the forensic biomechanist to determine if a mechanism was present to cause injury in a lowlevel fall.
Reliance on published scientific research by the forensic community should enhance the ability to accurately assess whether a mechanism of injury was present in a low-level fall. This presentation will
impact the forensic community and/or humanity by demonstrating that
caution must be exercised, however, when using published data to
perform the converse task, that is, to retrospectively predict injury from
a given fall. This is very complicated task dependent on many variables
including the large number of biomechanical factors affecting injury
potential and the wide range of injury types in the literature.
The impact energy available from a standing height fall is
derived from a conversion of the potential energy of the body’s
initial center of mass and can be sufficient to cause significant
injury, including fracture of the proximal femur, 1 distal radius, 2
lumbar vertebral body, 3 or skull; 4 but the majority of falls are
arrested safely without injury. Fall-related injuries occur when
the impact forces and moments exceed an individual’s biological
tissue tolerance to injury. DeGoede et al. 5 and Robinovitch et
al. 6 provide excellent review articles that document biomechanical factors contributing to injury severity in nonsyncopal
falls to the ground. This paper will not address all of these biomechanical factors; rather it will highlight some of those factors
that have experimental support in the scientific literature.
Direction of fall can influence both the primary location of impact
to the body in a fall and the subsequent injuries. Smeesters et al.7 found
that human subjects exposed to trips and unanticipated step-downs
during gait usually fell forward or sideways, whereas those exposed to
slips more often fell backward or sideways. Hsaio & Robinovitch8 analyzed body segment kinematics in young subjects during induced
forward, backward and sideways falls. In all falls, impact to the outstretched hand(s) was observed. All posterior falls also resulted in
contact with the pelvis. In greater than 70% of the falls with pelvis
contact, the time difference between hand and pelvis impact was less
than 50 ms. The investigators theorized that this short interval between
multiple impact points suggested a sharing of impact energy between the
upper extremities and pelvis which would likely reduce the injury risk
compared to a single impact point. The higher incidence of fracture seen
in the elderly population may be partly explained by slower upper
extremity reaction times or landing on the hip instead of the buttocks in
a low-level fall.
Extremity joint and soft tissue properties also govern extremity
impact force, energy absorption and injury potential. Chiu and
Robinovitch,2 using a mathematical model to simulate a forward fall
from standing height (0.75m) onto an outstretched hand, showed large
differences in the impact response between the wrist and shoulder. Peak
impact forces measured in the wrist (2.57 kN) surpassed the average
fracture force for the elderly distal radius (2.26.+ 1.01 kN), but these
116
large peaks were not transmitted proximally to the shoulder. The
shoulder underwent larger displacements and absorbed more energy
(277 J) than the wrist likely due to lower shoulder stiffness. This would
result in a higher potential for shoulder joint injury (e.g., rotator cuff tear
or capsular instability). Altering the extrinsic properties of the impact
surface with padding reduced the peak impact force at the wrist by about
35%,9 to a level that might prevent wrist injuries in standing height falls.
Unfortunately, padding did not generate a concomitant reduction in force
or deflection at the shoulder. A similar phenomenon has been observed
in experiments measuring peak head and neck loads following vertical
drops on the vertex of cadaveric heads with an average impact velocity
of 3.12 + 0.18 m/sec onto unpadded and padded surfaces.10 Padding
reduced peak force at the head but increased the impulse and catastrophic injuries in the cervical spine.
Volitional momentum arrest or energy dissipation strategies prior to
impact have also been identified in the scientific literature. Robinovitch
et al.11 showed that individuals have some control over the magnitude of
impact energy applied to their body during a low-level fall. In their
study these investigators simulated different reaction time delays to a fall
by asking standing human subjects to fall backward onto their buttocks
as softly as possible without use of their hands from 3 different initial
backward lean angles (Oº, 5º, and 12º). An increased initial lean angle
resulted in significantly increased vertical hip impact velocity and
kinetic energy at impact, and decreased energy-absorbing work at the
hip, knee, and ankle joints during descent. The increased lean reduced
the available reaction time and diminished the subjects’ ability to reduce
impact energy through a protective “squat” response that consisted of
lower extremity muscle contractions. The importance of reaction time
was also illustrated in research by van den Bogert et al.12 who used an
inverted pendulum model to show that faster response time was more
important than slower walking velocity for successful recovery from a
trip in older adults. The results of these studies highlight the importance
of intrinsic variables, such as reaction time, strength and flexibility, in
determining one’s ability to not only avoid falls but also dissipate energy
and affect impact severity in a low-level fall.
Anthropometry is a biomechanical variable that also influences
injury risk. Research has found a correlation between body mass index
(BMI) – weight (kg) divided by height2 (m) – and ankle fracture severity
in low-level falls.13 A BMI of 18 – 25 kg/m2 is considered desirable for
both men and women. In this study 24 patients with displaced malleolar
fractures were matched with 24 patients with undisplaced fractures. The
mean BMI of patients with displaced fractures (28.25 kg/m2) was
significantly higher than that (24.58 kg/m2) of those with undisplaced
fractures. These findings suggest that obesity is a risk factor for severe
ankle fractures in low-level falls.
Bone properties represent another biomechanical variable that contribute to the risk of injury from a low-level fall. Salminen et al.14 investigated 50 femoral shaft fractures from low-energy falls in a patient
population with an age range of 17-92 years. In 38 patients the fall
height was ground level. The significant biomechanical determinant of
fracture in this study was the presence in 64% of the patients of at least
one preexisting factor, besides age, likely to cause osteopenia, a
condition which weakens the mechanical strength of the femur.
In summary, reliance on published scientific research by the
forensic community should enhance the ability to accurately assess
whether a mechanism of injury was present in a low-level fall. Caution
must be exercised, however, when using published data to perform the
converse task, that is, to retrospectively predict injury from a given fall.
This is very complicated task dependent on many variables including the
large number of biomechanical factors affecting injury potential and the
wide range of injury types in the literature.
References:
1. Courtney AC, Wachtel, EF, Myers ER, et al. Age-related reductions in
the strength of the femur tested in a fall-loading configuration. J Bone
Joint Surg Am 1995; 77:387-395.
* Presenting Author
2. Chiu J, Robinovitch SN. Prediction of upper extremity impact forces
during falls on the outstretched hand. J Biomech 1998; 31:1169-1176.
3. McGill SM, Callaghan JP. Impact forces following the unexpected
removal of a chair while sitting. Acc Anal Prev 1999; 31:85-89.
4. Ono K. Current status of human head impact tolerance. In:
Yoganandon N, Pintar FA, Larson SJ, Sances A eds.
Frontiers
in Head & Neck Trauma. Amsterdam, Netherlands: IOS Press,
1998:183-199.
5. DeGoede KM, Ashton-Miller JA, Schultz AB. Fall-related upper body
injuries in the older adult: a review of the biomechanical issues. J
Biomech 2003;36:1043-1053.
6. Robinovitch SN, Hsiao ET, Sandler R et al. Prevention of falls and
fall-related fractures through biomechanics. Exercise and Sport Sciences
Reviews 2000; 28:74-79.
7. Smeesters C, Hayes WC, McMahon TA. Disturbance type and gait
speed affect fall direction and impact location. J Biomech 2001; 34:309317.
8. Hsaio ET, Robinovitch SN. Common protective movements govern
unexpected falls from standing height. J Biomech 1998; 31:1-9.
9. Robinovitch SN, Chiu J. Surface stiffness affects impact force during
a fall on the outstretched hand. J Ortho Res 1998; 16:309-313.
10. Nightingale RW, Richardson WJ, Myers BS. The effects of padded
surfaces on the risk for cervical spine injury. Spine 1997; 22:2380-2387.
11. Robinovitch SN, Sandler R, Torburn L et al. Impact severity during
backward fall depends on the timing of the “squat” protective response
during descent. Proceedings of 2001 Bioengineering Conference. 12.
Snowbird, UT: ASME, 2001:881-882.
van den Bogert AJ, Pavol MJ, Grabiner MD. Reponse time is more
important than walking speed for the ability of older adults to avoid a fall
after a trip. J Biomech 2002; 35:199-205.
13. Spaine LA, Bollen SR. The bigger they come…: the relationship
between body mass index and severity of ankle fractures. Injury 1996;
27:687-689.
14. Salminen S, Pihlajamaki H, Avikainen V et al. Specific features associated with femoral shaft fractures caused by low-energy trauma. J
Trauma 1997; 43:117-122.
Injury, Biomechanics, Falls
C12
Decoupling of Lagrangian Equations of
Motion to Improve Computational
Efficiency and Application to Multi-Body
Constrained Biomedical Engineering
Systems
Laura L. Liptai, PhD*, BioMedical Forensics, 1660 School Street,
Suite 103, Moraga, CA 94556; Bahram Ravani, PhD, University of
California at Davis, Davis, CA 95616
The goal of this presentation is to develop a unique method for formulation of equations of motion based on Lagrangian energy methods
that result in decoupling of the equations of motion in the highest order
terms. This derivation yields equations of motion that are in a more convenient and efficient form for numerical analysis.
humanity by demonstrating a method which reduces computer processing time by 10-17% which is significant when utilizing a high
capacity computer to analyze occupant motion. Computer processing
costs are also reduced proportionally.
As simulation of human occupant kinematics becomes increasingly
complex, computer processing effectiveness becomes an increasingly
important consideration. Decoupling the highest order terms improves
the efficiency of numerical analysis, especially for nonlinear systems.
More efficient processing yields results in less computer time or with
less computer processing capability. This method therefore, may widen
* Presenting Author
the practical use of occupant kinematic analytical tools.
Methodologically, the only work on decoupling equations of
motion by Loduha and Ravani (1995) utilized Kane’s method. Here, a
method for decoupling the equations of motion is derived for the first
time for use with Lagrangian energy based methods, commonly used in
BioMedical Engineering. Methodology involves mathematical transformation of the equations of motion. Specifically, the equations of motion
are decoupled in the highest order via a congruent transformation to a
diagonal matrix called the canonical form of a symmetric matrix under
congruent transformations. This was first attempted by Wade in 1951;
here, congruent transformation will be utilized for the first time to
decouple Lagrangian equations.
Highest order decoupling of Lagrangian equations of motion for
constrained multi-rigid body systems is formulated to create equations
of motion that are decoupled in the highest order terms. The practical
benefit is less computer processing time, which is most apparent for
complex systems. Decoupling the equations of motion in the highest
order derivative terms results in a more convenient form of the equations
for numerical analysis. Specifically, if the equations of motion are
decoupled in the highest order prior to integration, computer processing
is more efficient (especially for nonlinear systems) as decoupling of the
highest order is no longer required at each step.
Mathematical derivation requires seven steps:
1. Coordinate Selection
2. Lagrangian, Kinetic Coenergy, and Potential
Energy Transformations
3. Partial Derivative Transformations
4. Transformed Lagrangian Derivation
5. Transformation of Kinetic Coenergy
6. Transformation of Potential Energy
7. Selection of the Rate Transformation Matrix
In conclusion, a parsimonious side impact occupant kinematics
example demonstrates that the selection of generalized velocity components as a function of generalized coordinates indeed results in decoupling of the Lagrangian equations of motion in the highest order.
Further, a simplified BioMedical Model shows that the decoupled equations and the coupled equations of motion yield the same result. The
difference is the decoupled equations a require 10-17% less numerical
processing time.
Mathematical Modeling, Simulation, Human Kinematics
C13
Catastrophic Spinal Injury to Restrained
Occupants in Frontal Crashes
Carley C. Ward, PhD* and Hrire der Avanessian, PhD, Biodynamics
Engineering, Inc., 860 Via De La Paz, Suite B-3, Pacific Palisades,
CA 90272
Attendees will learn how restraint system design affects the
potential for catastrophic spinal injury through the analysis of fourteen
passive seatbelt cases.
humanity by demonstrating how certain seatbelt designs increase the
risk of catastrophic spinal injury in frontal crashes.
In this presentation, spinal injury mechanisms related to restraint
usage are identified and analyzed. The analysis of fourteen passive
seatbelt injury cases reveals how catastrophic spinal injuries are related
to the belt fit, torso belt load and belt geometry. Injury locations include
the lower cervical and upper thoracic spine. Although such injuries can
occur in high-speed impacts to a vulnerable person using an ordinary
three-point restraint, such injuries should not occur at lower speeds. Yet,
such injuries are not uncommon when automatic or passive belt systems
are utilized. Passive systems produce these injuries even in moderate
speed collisions. Some injury patterns related to single torso belts have
been reported. However, the reported injuries, their mechanisms andlo117
cations are, for the most part, different than those in this study. The
current study relates only to catastrophic spinal injury and to impacts
less violent than many reported in the literature. Even though these
restraint systems have been discontinued (passive belt restraints have
been replaced by airbags), analysis of passive belt injuries reveal dangerous design parameters that can be useful in evaluating risk factors in
other designs, such as the new all-belts-to-seat designs.
By definition, passive belt systems position themselves on the
occupant without any action by the occupant. The particular passive
design most frequently associated with these injuries is the passive torso
belt that travels on a track along the door or doorframe. This system has
a separate manual lap belt, but use of the lap belt does not prevent these
injuries.
The mechanism in all cases was neck flexion or flexion with distraction. The injuries were located at or between the C3 and T2 vertebrae. The outcome was paralysis from cord damage at those locations
or death. Women suffered the most injuries by a ratio of six to one. The
prevalence of women may be related to their typically shorter stature,
which can contribute to poor belt positioning, their lower neck strength,
and reduced shoulder-belt effectiveness because of the compliant female
chest. The age range was 17 to 84 years. But the age distribution indicates, as would be expected, that older individuals were more likely to
be injured.
Slightly more passengers were injured than drivers, but the difference was insignificant. The vehicles’ change of velocity ranged from
17 to 33 mph. A relationship between injury severity and change in
velocity was not apparent in this small data set. The shortest person was
4 foot 10 inches tall, while the tallest was 5 foot 10 inches. The mean
height was 5 foot 4 inches, or approximately the average height for a
woman. The weight ranged from 100 pounds to 260 pounds and no
obvious correlation with weight was found. Nine injuries involved C7;
this can be explained by the structural discontinuity that exists between
the cervical and thoracic spine at C7. The thoracic spine is less flexible
and buttressed by the ribs.
The factors common to these passive restraints that contribute to
injury are as follows. The torso belt track mount/anchor and inboard lap
belt buckle are mounted forward of the typical D-ring and buckle locations. The upper torso belt anchor is forward of the B-pillar, positioning
the torso belt forward of, and often several inches higher than, the
occupant’s shoulder. This geometry can position the belt dangerously
high against the neck of a short-statured occupant. The forward, upper
mount also created risk to taller occupants, who have their seats further
back. They have to move forward into the webbing before experiencing
any restraint. Thus, their chest and neck develop a velocity relative to the
belt before belt contact and they strike the belt with force. The reduced
length of the torso belt also increases the risk. This short length provides
less stretch. As a result, the belt develops high-tension forces.
Corresponding high stresses develop in the body under and adjacent to
the torso belt path, leaving obvious belt marks high on the upper chest
or at the base of the neck.
Contributing to the high torso belt loading and resulting injuries is
the forward position of the lap belt buckle. The inboard lap belt buckle
is forward and above the seat bite (intersection of seat back with seat
bottom) in some vehicles by up to seven inches. With this placement the
belt cannot fit snugly against the pelvis. Poor lower body restraint, due
to poor lap belt fit or no lap belt at all, allows the lower body to submarine relative to the torso belt. As a result, the torso belt moves up
towards the neck and distraction forces on the neck are increased.
Half of the injured individuals in this study were wearing their lap
belts. These occupants moved forward several inches before the lap belt
provided appreciable restraint. As a result, high torso belt forces
developed before the lap belt restraint became effective. Clearly the lap
belts included with these passive belt systems are ineffective in
restraining the lower body and distributing belt forces to the pelvis.
Failure of the lap belt to effectively restrain the lower body resulted in
higher shoulder belt forces than would be experienced with a continuous
118
loop three-point belt system. This was confirmed by MADYMO simulation of some cases.(1) MAthematical DYnamic MOdeling
(MADYMO) is a human body computer program for simulating threedimensional injury events. These simulations showed high torso belt
loads and the effect of the lap belt buckle location described above.
In summary, the injury mechanisms related to poor belt fit are identified. This study confirms the importance of good belt geometry, fit and
design. A restraint system without these qualities can cause unanticipated catastrophic spinal injuries, especially to women.
The authors wish to acknowledge Ann Mallory, Georgina Hackett,
Bruce Turpin and Caryn Doller for their work on these cases.
Reference:
(1) Ward CC and Der Avanessian H, “Passive Restraint Injury Analysis,”
TOPTEC, May 1995.
Passive Restraint Systems, Automatic Shoulder Belts,
Spinal Injury
C14
Head Impact by Golf Ball: Digital Data
Acquisition and Analysis Compared to
Alternative Methodologies
Moraga, CA 94556
quantify head accelerations and head injury criterion from being struck
in the head with a golf ball and compare these results to alternative
methodologies.
humanity by demonstrating the utility of three independent methods to
quantify head accelerations from impact from a golf ball.
This paper presents the analysis used to quantify the head accelerations and head injury criterion from being struck in the head by a golf
ball via three methods: digital data acquisition on a Hybrid III, scalp
tensile strength calculations and pressure sensitive film.
A 39-year-old female (height: 5’ 9”, weight: 145 lbs) was hosting a
league of 35 to 40 people on a Thursday evening at a golf course. After
greeting some of the new players, she proceeded to the putting green
where she was standing and looking down at a scorecard. She soon
noticed a painful sensation and became dizzy. There was no loss of consciousness. A gentleman from the ninth hole had driven his ball into the
trees and did not know where it went. A witness near the clubhouse saw
the ball bounce one time and then strike the golf instructor on the top of
her head. The instructor was escorted to the hospital by ambulance,
where they noted a small hematoma just right from the center-top of the
head. X-ray of the cervical spine was negative and brain CT revealed no
soft tissue swelling or discoloration.
Using distances measured from the golf course, angle of the golf
club face, golf ball lift and drag coefficients, and the equations of motion
it was determined that the maximum speed at which the golf ball would
have contacted the instructor was 27.5 mph. Using this impact speed, an
experiment was designed to drop a golf ball from an equivalent drop
height onto a Hybrid III dummy head to measure the resulting head
accelerations and the head injury criterion (HIC) value. In addition,
contact area was measured on the head form using pressure sensitive
film.
Methodologically, instrumentation and equipment used included a
tri-axial accelerometer mounted at the head center of gravity of a
Hybrid-III female anthropomorphic test dummy, compression 80 golf
balls, data filtering at 1650 Hz, and a data collection rate of 10,000
samples per second per channel.
The HIC became a US government standard in Federal Motor
Vehicle Safety Standard (FMVSS) 208 in 1972 and it is characterized by
the magnitude of linear head acceleration, duration, and indirectly,
impulse. Head injury in the form of concussion is predicted by a HIC
* Presenting Author
value greater than 1000. Six drop test trials were conducted with HIC
results of 1.3, 1.8, 1.4, 1.3, 2.0, and 1.7. It was found that the HIC was
below the threshold for concussion. HIC was utilized as a means to
quantify the accelerations and impulse to the head in this case; however,
HIC cannot be used as an injury threshold for all types of brain trauma
since one mechanical criterion could not adequately assess risk for all
types of head trauma diagnoses. Therefore, the use of HIC was limited
to the retroactive quantification of head trauma as in forensics and/or
comparative designs of head protection countermeasures.
Contact trauma analysis was to resolve if there should have been
tearing of the parietal scalp from the golf ball impact. Using the 27.5
mph impact speed, golf ball contact time, contact area, and mass, the
applied tensile load was calculated for the impact: 353.6 pounds/in2. The
ultimate tensile strength of the parietal scalp is 569.7 pounds/in2, which
would explain why there were no tears or lacerations in the parietal
scalp.
The HIC results of 1.3 to 2.0 over 0.5 milliseconds, in conjunction
with the lack of physical evidence of brain trauma, support the findings
that concussion would not be likely. This was supported by no definitive
loss of consciousness at the scene, no tearing of the parietal scalp and no
soft tissue swelling/discoloration.
HIC, Head Impact, Brain Trauma
C15
Accident Reconstruction of 14-Passenger
Catastrophic Rollover and Analysis of
How Occupant Restraints Could Have
Prevented Five Fatalities and Four
Serious/Severe Traumas Including
Analysis of Pediatric Restraint Usage
Suite 103, Moraga, CA 94556; Robert N. Anderson, PhD*, RNA
Consulting, 27820 Saddle Court, Los Altos Hills, CA 94022;
Michael E. Ensor, MD, JD, BioMedical Forensics, 1660 School Street,
Suite 103, Moraga, CA 94556
The purpose of this study wasto: 1) demonstrate the accident
reconstruction and tire integrity analysis involved in determining the
cause of this high speed solo rollover accident, 2) analyze seatbelt use,
3) assess the fatalities and serious/severe traumas that could have been
prevented with proper occupant restraint use, and 4) demonstrate a
unique approach to visual communication.
humanity by demonstrating how: 1) evidence can be utilized to support
findings that differed from other authority, 2) occupant trauma databases
were supplemented manually with the effect of proximity to crush, 3)
kinematic analysis of 14 occupants was simplified by utilizing a roofless
exemplar vehicle and 4) interactive presentation programs maximized
flexibility (individual occupants could be illustrated in any sequence or
in any level of detail).
History: Fourteen church volunteers, ages 5 – 70, were traveling in
a 15-seat passenger van. They were proceeding north on a rural
highway, a few minutes before 11 PM, when the rollover occurred. The
van was traveling at approximately 78 mph, and, due to some driver
impetus, it swerved to the right, initiating a right-side leading skid. This
led to a clockwise rollover.
Methods: First, the cause of the accident is determined using the
physical evidence revealed during the vehicle and scene inspections.
Second, the physical evidence in the vehicle and the trauma sustained by
the occupants is utilized to determine which of the 14 passengers were
restrained. The interior contents and the trajectories of those objects
were considered. Lastly, this information, in conjunction with: 1) crush
damage, 2) analysis of trajectories/ejection paths and 3) comparison to
databases1,2,3,4,5,6,7,8,9,10,11 of occupant exposure, is collectively utilized
* Presenting Author
to assess the trade-off injuries of those nine occupants that were determined to be unrestrained.
Cause of the Accident: Initially, in the case in question, there was
consideration that the front left tire was underinflated, causing steering
problems. Testing showed that the rollover could explain a loss of
pressure in the tire. Measurement of the roll angle versus lateral acceleration on an equaled loaded exemplar van followed the equation: roll
angle = 91.6(g)1.4. However, the rollover occurred on a relatively
straight section of a four-lane divided freeway that required little or no
steering input and had a maximum lateral acceleration of 0.07g for a roll
angle of 2.2 degrees. In fact, the driver caused the rollover by making
an unsafe turning movement while exceeding the speed limit and being
inattentive.
Restraint Usage and BioMedical Engineering Analysis: The van is
configured for 15 total persons with 15 occupant restraints. There are
driver and passenger captain’s chairs, three bench seats, each of which
with a three-person restrained capacity, and a last row bench seat with
four-person restrained capacity. In this case, the entire last row was
being utilized for storage and the front two seats were occupied, thereby
leaving four people in each of the three middle rows with only three seatbelts available in each row. The child was positioned sleeping over her
grandfather’s lap.
The subject vehicle was inspected to determine the amount of crush
sustained for each individual’s seating position and seatbelt use evidence. The seatbelt evidence reviewed in detail is the occupant’s
restraint use that differs from the MAIT report and the bases for the differences in opinion.
This analysis revealed that there were three properly belted and at
most two improperly belted passengers. Nine of the total fourteen occupants were ejected. Seven people died. Three occupants sustained
serious injury.
Next, resultant occupant clearances were determined for each
occupant’s position. The most severe crush was focused towards the left
front side of the vehicle.
The trade-off injury analysis utilized a database of collisions to
determine the most likely AIS levels, given the change in velocity of the
vehicle. The crush intrusion was greatest on the driver’s compartment
(who was seat belted properly). Given the severity of the intrusion, the
occupant would have perished regardless of seatbelt use. The four
people in the row behind the driver were unbelted and also perished,
most likely from their ejections from the vehicle. If properly belted and
seated correctly, only one of the passengers on that row at most would
have likely sustained life threatening injuries from the crush. Two
ejected passengers resulting in two fatalities from the middle row of
seats would likely have been avoided if the occupants had been properly
belted. The rest of the passengers would have sustained minor to moderate injuries if belted properly and seated correctly (i.e., using the last
row of seats) because there was little evidence of intrusion and force sustained to the rear occupants. Lastly, the pediatric out-of-position
occupant was analyzed.
Findings:
• the probable causes of the accident are: inattentive driver and
exceeding the 65 mph speed limit. There was no evidence of
mechanical failure, maintenance or road condition causal factors
for the rollover.
• Five fatalities and four serious/severe traumas may have been
prevented by choosing to use the seatbelts properly.
References:
1. Moffatt, E., Road Vehicle Rollover Collisions, University of
California, San Diego School of Medicine; presentation of “Accidental
Injury: Biomechanics & Prevention,” November 11-12, 1995, San
Diego, CA.
2. Otte, D., Südkamp, N., Appel, H., Variations of Injury Patterns of
Seat-Belt Users, SAE Paper 870226.
3. Huston, R.L., King, T.P., An Analytical Assessment of Three-Point
119
Restraints in Several Accident Considerations, SAE Paper 880398.
4. Grosch, L., katz, E., kassing, L., Marwitz, H., Zeidler, F., new
Measurement Methods to Assess the Improved Protection Potential of
Airbag Systems, SAE Paper 870333.
5. Viano, D.C., Biomechanics of Head Injury – Toward a Theory Linking
Head Dynamic Motion, Brain Tissue Deformation, and Neural Trauma,
SAE paper 881708.
6. Yoganandan, N., Pintar, F., Haffner, M., Jentzen, J., Malman, D.,
Wennshel, S.S., Larson, S.J., Nuchols, H., Sances Jr., A., Epidemiology
and Injury Biomechanics of Motor Vehicle Related Trauma to the
Human Spine, SAE Paper 892438.
7. Mackay, G.M., Parkin, S., Morris, A.P., Brown, R.N., The Urban
Rollover: Characteristics, Injuries, Seat-Belts, and Ejection, 1991 ESI:
Paper S6-O-08, Section 3: Technical Sessions.
8. Bahling, G.S., Bundorf, R.T., Moffat, E.A., Orlowski, k.F., The
Influence of Increased Roof Strength on Belted and Unbelted Dummies
in Rollover and Drop Tests, The Journal of Trauma: Injury, Infrection
and Critical Care, Vol. 38, No. 4, 1995.
9. Digges, K., Malliaris, A.C., Crashworthiness Safety Features in
Rollover Crashes, SAE Paper 982296.
10. Herbst, B., Forrest, S., Wang, P., Chang, D., Friedman, D., Friedman,
K., The Ability of 3 Point Safety Belts to Restrain Occupants in Rollover
Crashes, Paper Number 96-S5-O-12.
11. Biss, D.J., Safety Performance Evaluation of Slack Effects In ThreePoint Safety Belts Using the Hybrid III Dummy in Frontal and Frontal
Oblique Sled Tests, Presented at the sixteenth annual International
Workshop on Human Subjects for Biomedical Research.
including Barrier Equivalent Velocity, change of speed, direction of
impact, principal direction of force and impact duration, peak g versus
average g.
The result of each original concept is discussed and the new and
improved method of input or calculation follows. A very simple example
is to assume a set of four skids of different lengths. Assuming a coefficient of friction of 0.7 g, one takes the average length of the four skids,
according to a University Publication, to obtain the pre-braking speed.
A better method is to take the longest skid or to take the center of gravity
distance from start of skid to point of rest. However, this gives the speed
at the start of skid, not the start of braking. It is now recognized that 15
to 20 percent of the braking energy is expended prior to the visible skid
marks. This has to be added to obtain the probable cruise speed, which
is the speed at the start of braking. The old literature talks about the
coefficient of friction of the road. In fact, one needs to know the type of
vehicle, such as tractor-trailer combination, light truck, automobile, or
sports car, as well as the ambient condition for the particular road
surface. A more appropriate term than coefficient of friction is drag
factor which is measured in terms of units of g.
There are still states in America who are presenting 0.7 and 0.75
seconds for the daylight reaction time of a driver. The newer research
by Olsen and others and the accepted Perception Reaction time in most
State Superior Courts, the Criminal and Federal Courts is usually about
1.5 seconds.
The material in this paper has been shown to be enlightening to
even the seasoned forensic engineer, as well as stimulating and informative to the younger engineer.
Rollover, Seatbelt Analysis, Ejection
Older References, Reconstruction Updates, Reconstruction
Corrections
C16
Changes in Scientific Concepts in
Accident Reconstruction Since the 1960s
Philip V. Hight, BS, PE*, Accident Research and Analysis, 852 La
Milpita, Santa Barbara, CA 93105
After attending this presentation, the accident reconstruction specialist and forensic consultant will be conversant with changes in the
practice of accident reconstruction methods, constants, and inputs.
humanity by demonstrating an approach to correct misinformation in the
forensic arena. In addition, the younger research engineers in the
accident reconstruction field will find the development of the newer
research more meaningful.
The purpose of this paper is to help the accident reconstruction specialist become aware of older references and concepts that are often recognized but are in error and need to be updated. Most accident reconstruction is based on applied physics relying on Sir Isaac Newton’s three
principles. The proposition of this paper is to show that although the
basic physics is usually correct, the interpretation of input may give
incorrect answers. The method of updating the analytical calculations is
to review the latest research from the technical research papers presented
by the Stapp Car Crash Conference, the AAAM, Society of Automotive
Engineers, and the International Research Organization for the
Biomechanics of Injury, as well as technical papers from Canada,
Europe, and Australia, together with the research at the Trauma Research
Group, Dept of Surgery, UCLA. The questionable inputs and assumptions used for calculations have been printed in a number of accident
reconstruction publications and State and Federal brochures. The content
of this paper also includes the 1960s and 1970s research by the Trauma
Research Group, which showed that some of the input by Northwestern
University and many others in the accident reconstruction field was
incorrect. The areas and formulae covered in this paper include updated
concepts for the prediction of acceleration, cruise, cornering path, cornering acceleration, steering input, hydroplaning, perception reaction
time, emergency braking values, impending skid distance, crash events
120
C17
Air Bag Data: Useful for
More Than Air Bag Cases
Holly A. Adams, BSME*, ASA, Incorporated, 12015 Canter Lane,
Reston, VA 20191
use the data stored in the air bag computer to prove other facets of an
accident, such as sudden acceleration claims, whether or not cruise
control was activated, and identifying a spontaneous or inadvertent air
bag deployment.
humanity by providing a greater understanding of the information contained in automobile air bag computers.
THEORY OF THE ANALYSIS: As most of the participants are
aware, passenger vehicles equipped with air bags contain a computer as
part of the system that, depending on the make and model year, saves
various amounts of data. The earliest versions were simply diagnostic
units that monitored the air bag components, illuminated a light in the
dash and saved a fault code (DTC or diagnostic trouble code) if there
was a problem. This first generation also saved a history of the warning
lamp ON time. The next generation saves additional information relevant to the deployment of the air bags. This includes sensor closure
timing (which equates to air bag deployment timing) and the driver’s
seatbelt status.
The third generation of air bag computers saves Delta V (change in
velocity) or acceleration for the crash event. This technology employs
an accelerometer and a deployment algorithm to determine if the air bags
should deploy. Vehicles manufactured in the mid-1990s began to use
these systems, which are sometimes referred to as single point sensing
systems because many of them eliminated forward crush sensors. By the
21st century, this third generation of sensing systems/software has been
upgraded to include pre-crash data (GM only). Pre-crash data is a record
of the vehicle’s speed (both mph and rpm), throttle application and
braking (on or off) for a predetermined amount of time before the crash
* Presenting Author
event. This data has become extremely useful to reconstructionists to
verify their analysis of the accident sequence.
With the advent of more sophisticated safety technologies, such as
side air bags, dual stage air bags and occupant sensing, more information
will become available from the air bag computer. Items such as the front
passenger’s seatbelt status and the front seat occupant’s proximity to the
air bag will be saved in order to allow the air bag computer to make the
decision whether or not to deploy the air bags. This will also mean that
having only one frontal air bag deploy in a crash is not necessarily an
automatic defect.
Several case studies will be presented in which the air bag computer
contained information helpful to a case in which the air bag was not necessarily the primary focus.
Case 1 involved an elderly driver in a parking lot of a post office.
The subject vehicle went out of control, through the post office window
and pinned another elderly person to the wall, causing fatal injuries to
the postal customer. The driver insisted that the vehicle took off and,
despite pressing hard on the brake, could not be stopped. The data stored
in the subject vehicle proved otherwise.
Case 2 involved a vehicle that became disabled on the highway in
a travel lane with no electrical power, hence, no hazard lights. The
driver of the disabled vehicle got out of the vehicle and went to the
shoulder of the road. The driver witnessed her vehicle get side-swiped
twice. A third vehicle (an SUV) came up behind her disabled vehicle
and swerved onto the shoulder instead of the other lane. The SUV hit
the driver of the vehicle and killed her. The driver of the SUV swore he
was going the speed limit and that the cruise control was off. The data
stored in the subject vehicle proved otherwise.
Case 3 involved a vehicle driving down a well-traveled dirt and
gravel road. The driver claimed that the air bags just went off, causing
a loss of control, but ultimately no subsequent accident. The lack of
damage to the vehicle and the data stored in the subject vehicle proved
that the driver was telling the truth.
Air Bag, Data, Interpretation
C18
Road Rage Caused a
Multiple Vehicle Crash
Donn N. Peterson, MSME*, Peterson Engineering, Inc., 15720 15th
Place North, PO Box 47565, Plymouth, MN 55447-0565
application of engineering analyses to determine a crash sequence
leading to the conclusion that the cause was road rage of a driver whose
vehicle was the last one to become involved in any of the multiple collisions.
humanity by demonstrating a basis for accident reconstruction in
complex collisions. Elementary calculations may give part of the story,
but not necessarily a complete solution.
Background: On a summer night Ms. BB was driving a rented van
truck loaded with household furnishings and towing a low bed trailer
loaded with a small sedan. She was westbound on I-80 in central Iowa
with her son AM as a passenger when she passed a westbound loaded
semi truck driven by Mr. DR. A few minutes later and a few miles down
the road, Mr. DR passed Ms. BB such that his semi truck was westbound
on I-80, generally in front of Ms. BB, who was now in the westbound
lane. Mr. MG was driving his luxury sedan with front seat passenger AC
westbound on I-80 following Ms. BB. Mr. KP was driving a second
loaded semi truck westbound on I-80 in the right lane, following Mr.
MG. Shortly after midnight, the front of Mr. KP’s semi truck collided
with the rear of Mr. MG’s luxury sedan. Mr. MG’s luxury sedan rotated
½ revolution and its rear end collided with the rear of the trailer and the
small sedan being towed by Ms. BB. Mr. KP’s semi truck continued
forward and its front right collided with the left rear of Mr. DR’s trailer.
* Presenting Author
Ms. BB’s van truck came to rest facing WSW in the north ditch, a
short distance beyond the beginning of a weigh station exit ramp. Her
trailer with the loaded small sedan separated from the van truck hitch
and came to rest facing WSW in the north ditch with its left side near the
van truck right side. Mr. MG’s luxury sedan came to rest facing ENE in
the north ditch a short distance east from Ms. BB’s van truck and trailer.
Rear portions of Mr. MG’s luxury sedan were damaged by fire after its
two collisions. Mr. KP’s semi truck came to rest facing W and straddling
the dividing line between the two westbound driving lanes with its trailer
partly jack-knifed SW. The rear end of Mr. KP’s trailer was a short distance SSW from the front of Ms. BB’s van truck. Mr. DR’s semi truck
came to rest on the weigh station exit ramp facing WNW with the rear
of his trailer a short distance due south from the front of Ms. BB’s van
truck.
Another investigator photographed the accident scene and vehicles
within a day or two after the crashes. A few days later, with the investigator’s photographs in hand, the author inspected the scene and Mr.
MG’s luxury sedan. Pertinent measurements were made.
Application of engineering and crash analyses and accident reconstruction methods will be described, and the following results will be
presented:
• Mr. KP’s semi truck speed was over the 65 mph maximum speed
limit. He was in the right part of the right westbound lane turned
to the left with brakes applied hard.
• Mr. MG’s luxury sedan speed was under the 40 mph minimum
speed limit. He was in the right westbound lane near the north
shoulder pointed to the right and steering to the left. The space
between his vehicle and Ms. BB’s trailer was much less than a
safe operating distance.
• Ms. BB’s van truck speed was under the 40 mph minimum speed
limit. The van truck was straddling the north fog line between the
right driving lane and the north shoulder, and the trailer was in the
right driving lane near the fog line. The van truck and trailer were
turned to the right toward the north ditch.
• Two different scenarios were explored for Mr. DR’s semi truck
actions. In both scenarios his speed was much slower than the
40 mph minimum speed limit, impeding a normal traffic flow. In
one scenario he is moving very slowly (less than 15 mph) in the
right hand lane before turning onto the weigh scale exit ramp. In
the other scenario he is moving to the right at a slightly faster
speed and is literally running Ms. BB off the road. In either case
he is grossly interfering with Ms. BB’s travel path. (Her son and
passenger AM stated that he thought Ms. BB had passed Mr. DR’s
semi truck several miles before the collisions began.)
Crash Analyses, Accident Reconstruction, Road Rage
C19
Air Bag Control Module Response
to Multiple Vehicle Impacts
Kurt D. Weiss, BSMS, MSME*, Automotive Safety Research, Inc.,
5350 Hollister Avenue, Suite D, Santa Barbara, CA 93111-2326; Holly
A. Adams, BSME, Automotive Safety Analysis, 12015 Cantor Lane,
Reston, VA 20191
After attending this presentation, attendees will understand how the
timing between impacts, even minor ones, can explain the nondeployment of a frontal airbag in an event that is clearly above the
deployment threshold.
humanity by demonstrating the analysis of timing between multiple
vehicle impacts, and the ability of airbag control modules to adequately
protect vehicle occupants in these common situations.
THEORY OF THE ANALYSIS: The primary function of an airbag
control module is to predict impact severity, and to deploy the airbags
when necessary. However, when vehicles are exposed to two impacts in
121
close succession, the airbag control module may not be able to recognize
the more severe second impact. Without the benefit of a supplemental
inflatable restraint, the seat belted driver can sustain significant head and
facial injuries. This paper discusses the phasing of two impacts during
which the vehicle’s airbags did not deploy.
Prior to a tree impact, a vehicle veered sharply to the right and
struck a curb at the roadway edge. The vehicle’s momentum was sufficient enough for front wheels to rise over the curb. After the curb impact,
the vehicle was involved in a more severe second collision with a tree
having a diameter of approximately 15 inches. This tree impact caused
considerable property damage to the front centerline of the vehicle. As
a result of the tree impact, the driver’s head forcefully struck the steering
wheel rim. The steering wheel rim was deformed, and both shear capsules were completely separated. The driver sustained a severe laceration to the forehead requiring reconstructive surgery. Witness statements supported by physical evidence on the seat belt system indicate
the driver was wearing the available 3-point lap and shoulder belt at the
time of collision. However, the airbags did not deploy.
Collision data retrieved from the vehicle’s Sensing and Diagnostic
Module (SDM) indicated a pre-impact speed of approximately 22 miles
per hour. A near deployment event was recorded by the SDM with a
maximum velocity change of 0.21 miles per hour at 35 milliseconds
after the sensing algorithm was activated (AE). A near deployment event
is an impact that is not severe enough to warrant activation of the
airbags, but initiates the computer’s crash discrimination algorithm.
This near deployment event is consistent with the collision severity of a
curb strike.
A geometric analysis of the roadside dimensions was undertaken.
Based on vehicle specifications, the relative approach angle to the curb,
and the location of the tree, the vehicle traveled approximately 5.4 to 7.9
feet between the curb and tree impacts. The time between these two
impacts was approximately 168 to 245 milliseconds. Based on the
recorded data, the SDM was actively analyzing the crash for 40 ms.
Therefore, there was only approximately 128 to 205 milliseconds of time
for the SDM to record, reset and resume collision detection before
impact with the tree.
Based on published specifications of microprocessors of the type in
this vehicle, it takes approximately 10 ms to write a byte of data, and the
time required for system reset is approximately 200 to 500 milliseconds.
There are a total of 80 bytes of data stored in a near deployment record,
of which 56 contained a value other than $FF (the unwritten value).
Therefore, the vehicle struck the tree when the SDM was continuing to
reset. As a result, the SDM was unable to evaluate the second collision
within adequate time to command the deployment of the airbags. The
injuries sustained by the driver in this impact would have been significantly reduced by the added protection of an airbag.
Air Bag, Multiple Impacts, Timing
C20
Forensic Testing of Shared
Anchor Seat Belt Components
Kurt D. Weiss, BSMS, MSME*, and William G. Broadhead, MSME,
Automotive Safety Research, Inc., 5350 Hollister Avenue, Suite D,
Santa Barbara, CA 93111-2326
After attending this presentation, attendees will understand how
inadequately designed seat belt components, while saving production
costs, can significantly reduce the ultimate strength of the assembly in
real world traffic accidents.
humanity by demonstrating how inadequately designed restraint systems
can fail in real world traffic accidents.
THEORY OF THE ANALYSIS: Seat belts remain the primary
safety device in reducing the risk of ejection in motor vehicle collisions.
Ejection increases the risk of serious injury by a factor of 4.5. Pursuant
122
to federal safety standards, hardware for the type 2 upper torso restraint
shall be designed and located in the seat belt assembly such that the possibility of injury to the occupant is minimized. However, in real world
accidents, it has been observed that some seat belt designs can be inadequate. Failure of seat belts can render vehicle occupants unrestrained,
thereby increasing the risk of serious injury and ejection in rollover collisions.
A passenger van with seven occupants was traveling at highway
speeds. Suddenly, an impact by a vehicle traveling in the same direction
sent the van out of control. The van yawed clockwise, rolled over multiple times, and came to rest in the shoulder. Two rear seat occupants
were ejected, one suffering severe head and chest injuries.
Inspection of the two adjacent seat belts revealed physical evidence
confirming that the rear occupants were wearing their lap and shoulder
seat belts at the time of collision. It was found that both seat belt buckles
were designed to be secured to the seat frame by a single webbing strap
passing through one bracket. However, it was determined that the
stitching used to assemble the component failed, releasing the buckles
from their anchorage.
Documentation of the history of this shared anchor component
showed there was a design change during the vehicle production run.
The former component design used two similar buckles, each attached to
an anchor bracket with independent webbing straps. With this design,
occupant loading was transferred through the webbing strap to the
bracket attached to the seat frame. The redesigned buckle component
uses one piece of webbing to attach two buckles to one anchor bracket.
The webbing is routed through the two buckles and bracket, and the
layers of webbing are stitched together. Documents show this redesigned
component produced a 10¢ per end item savings, or a 24¢ per vehicle
savings. However, the ultimate strength of the redesigned component is
decreased by virtue of the stitching configuration used to assemble the
component.
A series of twelve tension tests was conducted on new and used
samples to evaluate the effect of loading angles on the failure strength of
these two shared anchor component designs. The loading angle is
defined as the included angle between the force vectors directed along
the individual webbing straps of the adjacent seat belts. Three samples
each were tested at loading angles of 60, 90, 120, and 180 degrees.
A rigid beam fixture was fabricated to establish the webbing strap
angles of 60, 90, and 120 degrees. The buckle component was secured
to the test bench using the factory original anchor bracket. To apply
tension to the component, a latch plate was inserted into each buckle,
and webbing routed through the latch plate was held by a split-drum grip
as specified under FMVSS 209. The rigid beam was attached to the
crosshead of an Instron tension-compression machine, and raised at a
constant rate of 100 millimeters per minute.
The rigid beam was not used for the 180 degree loading angle. For
this loading angle, a latch plate was inserted into each buckle, and
webbing routed through the latch plate was held by split-drum grips.
One split-drum grip was secured to the test bench. The other web grip
was attached to the crosshead, and raised at a constant rate of 100 millimeters per minute.
At a 60 degree loading angle, the former component design did not
fail during the test, while the redesigned component failed at an average
force of 11,899 Newtons. At a 90 degree loading angle, the former component design did not fail, while the redesigned component failed at an
average force of 6,592 Newtons. At 120 degrees, the former component
design did not fail, while the redesigned component failed at an average
force of 4,467 Newtons. Lastly, at a loading angle of 180 degrees, the
former component design failed at a force of 22,153 Newtons, whereas
the redesigned component failed at an average force of 2,169 Newtons.
Component designs can significantly effect the strength of seat belt
attachments. The test series established that some designs reduce the
strength of the seat belt so much they no longer satisfy the FMVSS 209
requirement for webbing breaking strength. Inadequate designs can
result in restraint failure when the shared anchor component of adjacent
* Presenting Author
seat belts are loaded with reasonably anticipated forces. Designers and
manufacturers must consider failure mode effects analyses before implementing a component design change in order to reduce production costs.
Seat Belt, Anchorage, Testing
C21
Evaluating the Effect of Assembly
Hardware on Breaking Strength
of Seat Belt Webbing
Kurt D. Weiss, BSMS, MSME*, Automotive Safety Research, Inc.,
5350 Hollister Avenue, Suite D, Santa Barbara, CA 93111-2326
effect of seat belt component designs in relation to reducing the ultimate
strength of the seat belt webbing.
humanity by demonstrating seat belt designs which are certified pursuant to federal safety standards can fail at force levels below the
intended required values.
The injury-mitigating potential of seat belts in motor vehicles is
widely accepted. Seat belts must be certified to conform to all
applicable federal safety standards. However, in real world loading conditions, it has been observed that some seat belt assembly hardware can
reduce the breaking strength of the webbing, apparently to levels below
those specified in Federal Motor Vehicle Safety Standard (FMVSS) 209.
Two test series were conducted on new and used seat belt assemblies to
evaluate the effect of assembly hardware on the breaking strength of seat
belt webbing under reasonably anticipated forces due to occupant
loading.
The first series contained four tests to evaluate the breaking
strength of the webbing alone, without any effect from the latch plate/
buckle component. The webbing samples tested were sectioned from
the portion of webbing adjacent to the seat belt retractor. Each end of
the webbing was held by split drum grips as specified under FMVSS
209. One split drum grip was rigidly anchored to the test bench, while
the other grip was accelerated by a hydraulic cylinder to a speed of
between approximately 0.91 and 1.22 meters per second.
To investigate the effect of the latch plate design on reducing the
breaking strength of the webbing, a second series of tests was performed.
The second series contained sixteen tests on sixteen different webbing
samples. Three-point anchoring held the seat belt webbing in a “V”
shape, with an included angle of approximately 45 degrees, similar to invehicle use geometry. One end of the webbing was secured to a
hydraulic cylinder using the OEM anchor bracket to which the webbing
was sewn. The hydraulic cylinder provided a pelvic restraint webbing
pre-load between 227 and 454 kilograms. The end of the upper torso
restraint webbing was held by a split drum grip. The split drum grip was
accelerated by a hydraulic cylinder to a speed of approximately 1.5
meters per second.
FMVSS 209 specifies that the webbing of a type 2 seat belt
assembly shall have a breaking strength of not less than 2,270 kilograms
for the pelvic restraint, and not less than 1,810 kilograms for the upper
torso restraint. The first test series showed that without stress concentrations induced by the latch plate, webbing failure loads ranged between
1,887 and 2,209 kilograms. When compared to the requirement of
FMVSS 209, these loads are somewhat less than the 2,270 kilograms
minimum strength for the pelvic restraint.
FMVSS 209 specifies that any webbing cut by the hardware during
test shall have a breaking strength of not less than 1,590 kilograms at a
cut in webbing of the pelvic restraint, or not less than 1,270 kilograms at
a cut in webbing of the upper torso restraint. The second test series
demonstrated that the webbing was being cut by the seat belt buckle
hardware. The metal edge of the locking latch plate caused stress concentrations and allowed the webbing to rip much like a ruler is used to
* Presenting Author
tear paper. This metal edge is common to both the pelvic and the upper
torso restraint. The measured loads at failure range from 827 kilograms
to 1,103 kilograms, and are far below those specified by the standard.
The test series demonstrate that the breaking strength of the seat
belt webbing material is dramatically reduced by the locking bar of the
latch plate assembly. Some latch plate designs reduce the strength of the
seat belt system so that they no longer pass the intent of the applicable
Federal Motor Vehicle Safety Standards. This reduction in restraint load
capability results in a vulnerability to total loss of seat belt restraint in
traffic collisions.
Seatbelt, Failure, Testing
C22
Vehicle Steering Caused by Tire Blowout
Grahme Fischer, MS*, Technical Problem Solvers, Inc., 900 Wheeler
Road, Hauppauge, NY 11788
The purpose of this presentation is to provide the forensic community with an analysis of an unusual automotive accident in which a
hole in a tire sidewall caused an abrupt turning maneuver.
humanity by demonstrating how a seemingly small defect can have very
large consequences.
This paper presents an analysis of the effects of a left-front tire
blowout on a 2-day-old 1990 Nissan Pathfinder. The subject vehicle suddenly turned left and struck an oncoming vehicle on a narrow two-lane
roadway. Immediately after the collision, the responding police officer
discovered a hole in the outboard sidewall of the Pathfinder’s left front
tire. A major issue of the resulting liability case was whether the accident
was due to the Nissan driver’s error or was caused by the tire failure.
This paper will show that the accident was caused by the tire failure.
Specifically, the paper will present evidence that:
• The tire’s sidewall hole was not caused by the collision;
• If the hole was present before the collision, the force of escaping
air from the hole would have caused the vehicle’s front wheels to
rapidly steer left; and
• The vehicle did steer left.
The subject tire was examined. It was determined that the sidewall
hole was created from the inside of the tire and not by some external
cutting object. That conclusion made it likely that the hole was created
before the collision, by the following logic: Engineering judgment
(unquantified) suggests that the probability was near zero that such an
unusual tire failure could occur in a collision event during which the
driver also performed an inexplicable steer-left maneuver into oncoming
traffic. That is, two extremely improbable events did not occur during
the same collision. Based on the following analysis of forces that would
have been exerted on the tire by escaping air, it was concluded that the
hole appeared in the tire’s sidewall and caused the rapid left-turn
maneuver into oncoming traffic, which caused the subject collision.
When the hole was created in the tire’s sidewall, high-pressure air
within the tire escaped through the hole into the atmosphere. Maximum
flow rate was achieved when air velocity reached the speed of sound, a
condition known as choked flow. Choked flow lasted for about one half
second after the sidewall rupture, lowering both the source and critical
pressures until the flow was no longer choked. During this time of
choked flow, the escaping air developed a force perpendicular to the
sidewall of the tire that ranged from 22.5 pounds to 11.7 pounds, creating an average thrust force of 17 pounds. At the Pathfinder’s estimated
speed of 30 mph, the tire (and the hole) made 2.7 rotations during the
half second of choked flow.
A total-station surveying instrument was used to measure the
spatial relationships among tire footprint, wheel spin axis, and wheel
steering axis. As the vehicle moved forward and the left front tire rotated
about its spin axis, the force of the escaping air produced a torque on the
steering axis. When this force (i.e., the hole) was forward of the wheel’s
123
steering axis it produced a clockwise (CW) torque (as seen from above)
which tended to steer the wheel into a right turn. Similarly, when the
force from escaping air was aft of the steering axis, it produced a
counter-clockwise torque (CCW) tending to steer the wheel into a left
turn. On the subject vehicle, the steering axis was just forward of the
wheel’s spin axis at the axle, so as the jet of air revolved about the axle
it spent a greater amount of time behind the steering axis than in front of
it, and the torque-arm from hole to steering axis was larger when the hole
was behind the steering axis than when the hole was in front of it.
Therefore, the CCW (leftward) steering torque produced by the hole
lasted longer and was stronger than the CW (rightward) steering torque
as the hole traveled one revolution.
The steering motion of the front wheels under these conditions was
dependent on the rotational position of the tire when the hole first
appeared. For this analysis, it was assumed that the hole appeared when
the tire’s sidewall was under maximum stress, i.e., when the hole would
have been at the six o’clock position.
In order to quantify the left-steer theory, steering torque was calculated and plotted against time for ½ second, starting with the hole at the
six o’clock position. The result was the decreasing-amplitude sine wave
shown in Figure 1. Integration of the steering torque versus time curve
produced the steering impulse curve shown in Figure 2. The impulse
varied in amplitude but always increased a left-turn steer angle. Thus,
the wheel always turned to the left during the time of choked flow.
There were no data available on the resisting torques of tire footprints to steering rotation. Common driving experience in the era before
power steering revealed very large resisting torques at very slow or zero
forward speed, and extremely low resisting torques at higher speed. For
simplicity, and recognizing the existence of inaccuracies in this method,
analysis of vehicle response to sudden discharge of air through the
sidewall assumed zero resisting torque at the tire footprint and from the
steering linkage.
This paper will supply the following evidence in support of the
above argument:
• Photographic evidence demonstrating that the tire failure
originated on the inside of the tire’s sidewall;
• An explanation of why the hole in the tire’s sidewall could not
have been created during the collision;
• An analysis of the forces developed by the airflow through the tire
sidewall;
• An analysis and graphical results of the steering torque produced
by airflow out of the hole in the tire;
• Graphical results of an analysis of estimated steering wheel
position versus time; and
• A representation of vehicle behavior in response to the
assumptions of the above analyses.
The subject accident caused substantial emotional turmoil (and
physical injury) to the driver and passenger of the Pathfinder. The driver was
very relieved to learn the above explanation of the accident. He had been
blaming himself for over 10 years for the collision and its resulting harm,
even though he knew that he had not steered left into oncoming traffic.
Steering, Tire Blowout, Accident Reconstruction
C23
Trailer Loading Ramp Defects Cause
Scissors Lift to Tip When Unloading
Donn N. Peterson, MSME*, Peterson Engineering, Inc., 15720 15th
Place North, Plymouth, MN 55447-0565
humanity by demonstrating analytical methods and simulation techniques which are useful in understanding phenomena and demonstrating
relevant events. They are also effective in testing hypotheses when
physical tests or reenactments are not feasible.
Background: A self-employed drywall applicator rented a narrow
slab scissors lift for a commercial building job on which he was
working. When his need for the scissors lift was finished, he borrowed
a flat deck trailer and pickup truck from his business partner to return the
scissors lift to the rental dealership. He loaded the scissors lift onto the
trailer and drove to the rental dealer accompanied by his friend as his
passenger. When he arrived at his destination, he parked the pickup and
trailer on the asphalt paved street in front of the rental dealer’s establishment. He manually placed the portable loading ramps into position
at the rear of the trailer, climbed to the operator’s station of the scissors
lift, and backed the scissors lift from the trailer onto the ramps. His
friend was standing on the street behind the trailer near the left side ramp
helping to guide him during this maneuver. While he was backing down
the ramps, the left ramp dislodged from its trailer support and the
scissors lift tipped onto its left side. He managed to jump from the
operator’s station to safety. His friend’s right foot was pinned and badly
crushed between the left side of the scissors lift and the street.
The subject trailer, loading ramps, and pickup truck were inspected,
measured, and photographed at the rural residence of their owner. The
subject scissors lift was inspected, measured, and photographed at the
rental dealership. The incident scene was inspected, measured, and photographed. Since there was less than full cooperation from the owners
of the equipment, the various pieces of equipment were not brought
together at the incident site. It was judged to be infeasible, and probably
too dangerous, to perform a physical reenactment of the incident.
Analytical methods and simulation techniques provide an alternative to physical testing for understanding and demonstrating relevant
events leading to tip over. Several key frame “building blocks” used in
constructing animations can be derived and demonstrated. Since the
elapsed times between these key frames cannot be accurately and
reliably determined from the known data, scientific animations cannot
be produced to demonstrate motions of the equipment. However, these
key frames can be used to demonstrate sequences of events in a stopaction series of still images format.
AUTOCAD 2000 was used to generate 3-dimensional scale
drawings and images of relevant features of the equipment involved.
These images were combined adhering to applicable physical constraints. Early during the trailer and ramp inspection, it was noted that
the tops of the ramps rest in a shallow slot on the rear of the trailer and
are held in place by gravity with no locking device. The mechanism of
dislodgement became apparent only after detailed engineering analyses
as demonstrated by the key frames.
The bottom ends of the ramps are shaped such that the ground
support point is actually 6 ½ inches forward from the rear edge of the
ramp platform. When a sufficiently heavy load is placed on the rear end
of the ramp, this permits the ramp to pivot lifting the front end of the
ramp from its support slot on the trailer. A small horizontal movement
of the ramp front end will then cause the ramp to miss its slot and fall to
the ground when the load is removed. The subject scissors lift wheelbase
is 12 ½ inches longer than the length of the ramps; this was a significant
feature in causing dislodgement of the ramp during unloading.
Loading Ramp, Design Defect, Tipping Stability
application of analytical methods and simulation techniques in testing
hypotheses and demonstrating a physical event where physical tests and
reenactments are not feasible. Key frames can be used to demonstrate a
sequence of events in a stop-action series of still images even when a scientific animation cannot be produced.
124
* Presenting Author
C24
Modern Equipment Control
Affects Electrocution Risk
John A. Talbott, BS, PE, Talbott & Associates, Inc., 7 SE 97th Avenue,
Portland, OR 97216-2498; Louis F. Bilancia, BSEE, PE*, PO Box
68103, Oak Grove, OR 97268
After attending this presentation, attendees will be able to identify
some work place electrical hazards, to expose the elements of electrocution, and provide a comparison of SCR switching and rheostat control
to an electrocution.
humanity by demonstrating how to identify some electrical hazards in the
workplace, and how the use of modern electrical design techniques may
contribute to these hazards.
Proposition/Hypothesis: A fatal industrial accident involving a hotwire foam cutter indicates a potential increase in electrocution hazard
resulting from the use of modern switched-mode techniques in industrial
control design.
A salesperson at a foamed plastic plant was using a hot wire cutter to
cut to size a stack of three 2” thick panels of structural foamed plastic. It
was an extra order which the person was performing alone on a very hot
night. He had been working for 17 hours when he was found positioned
as though he had tumbled awkwardly into the frame of the cutting table.
The coroner reported a cauterized lateral burn on the decedent’s face, with
charring on the interior of lip and a superficial burn on the thigh corresponding to the frame height. Cause of death was noted as positional
asphyxia.
The knob controlling cutting wire heat was found rotated to the zero
position; the power switch was on, but the switches controlling cutting
wire carriage movement were off.
With the controls in this position, the potential from the cutting wires
to ground was measured to be 120 v.
It was surmised that the decedent was sighting the cutting wires to
position the work piece when he contacted the topmost of three cutting
wires with his sweaty face. This sent an electric current from his head
though his neck and torso to his right thigh.
Measurement of the electrical resistance of the body of other subjects
demonstrated that an electrical impulse sufficient to affect the heart would
have been generated by that contact.
Muscles are electrochemical in nature and, as such, produce characteristic electrical waveforms but can also be influenced and controlled by
electrical signals. Muscles and nerves generally reside in a polarized state,
poised for action. Once a muscle or a nerve is activated it “depolarizes”
thus transmitting the intended signal (for a nerve) or performing a physical
contraction (for a muscle). When they recover, by redistributing metallic
ions in the body-fluid electrolytes, they are said to repolarize.
Electricity follows the path of least resistance. In living tissue the
paths of least resistance are the nervous system, the lymphatic system, and
the circulatory system. With regard to the heart and the cardiac cycle,
there is significant sensitivity at one particular place in the cardiac cycle
referred to as the T wave. The normal cardiac cycle exhibits an atrial
depolarization demarking the contraction of the right atrium; this is identified as the P wave. After a variable 125-300 millisecond delay, another
depolarization referred to as the QRS-complex, demarks the contraction of
the ventricles of the heart. Following the QRS complex by 50 to 150 milliseconds is the T wave, which demarks the repolarization of the cardiac
muscle.
Modern defibrillator technology uses IGBT, Insulated Gate Bipolar
Transistor, similar to the SCRs used in industrial control, in switchedmode controlled circuitry to produce a stream of pulses between 50 and 60
Hertz to perform cardiac conversion. The term “conversion” in the
context of cardiology refers to converting a chaotic or fibrillating rhythm
to a regular pacing rhythm. The same stream of pulses can convert a
regular cardiac rhythm into fibrillation or from fibrillation to a regular
rhythm.
* Presenting Author
Modern semiconductor components have increased in performance
such that with modern pulse-width and frequency-modulation control
techniques, the use of rheostats, iron core inductors and transformers
which limit the controlled circuit voltage is being supplanted. Such techniques are good in that they decrease the size, weight, and cost of industrial controls; however, the control waveforms bear a significant resemblance to the waveforms intentionally used by cardiac defibrillators.
In conclusion, there is a particular electrical hazard present in the
modern industrial environment due to the use of the modern electronic
design techniques afforded by improvements in silicon devices.
Other factors in this case are (1) failure to turn off the machine’s
main switch; (2) two red warning lights were burned out; (3) required
warning signs were absent or obliterated; (4) decedent was doing work
he was told not to do; (5) decedent was probably affected by the long
hours and high heat; (6) decedent was not a regular operator of the
machine.
Electrocution, Cardiac, Switched-Mode Control
C25
Electrical Accident, Sabotage, Homicide,
or Death by Other Means?
Helmut G Brosz, BASc, PEng, Brosz and Associates, 64 Bullock Drive,
Markham, Ontario, Canada
At this presentation, a case will be presented in which electrocution
was speculated to be the possible manner of death. Investigation will
reveal whether it was an electrical accident, sabotage, homicide, or death
by some other means.
When the Olympics came to Atlanta in the summer of 1996, the city
was in a “feel-good frenzy” as a writer in The Atlanta Journal
Constitution put it. Hundreds of thousands of people, both locals and
out-of-towners, were enjoying the “high-octane mixture of athletic
excellence and world-class partying.”
The Olympic games where held at Centennial Olympic Park, which
was constructed specifically for the Olympics but remains a permanent
structure of downtown Atlanta.
On Friday, July 26, the ninth day of the 1996 Summer Games, a
pipe bomb placed near the main stage in the park exploded, injuring
more than 100 people, many of them permanently, and killed Alice
Hawthorne, a mother who had traveled to Atlanta with her daughter to
see the Olympics. The fatal bombing in Atlanta was a terrorist attack
aimed at thousands of innocent persons gathered at the Olympic Park.
Eric Rudolph has been charged with the bombing.
He is also accused of the January, 1997, double bombing outside a
suburban Atlanta clinic that performed abortions, the double bombing of
an Atlanta lesbian nightclub a month later, and the January, 1998,
bombing of a Birmingham, Alabama, clinic that performs abortions.
On April 18, 2000, an FBI Agent was suspected to have been
fatality electrocuted by means of a sabotaged appliance in his cabin residence in Andrews, North Carolina by Eric Rudolph. The agent was on
assignment in searching for Rudolph who is believed to have gone into
hiding after the last bombing in the 517,000-acre Nantahala National
Forest (also known as the Smoky Mountains) in western North Carolina.
The FBI engaged this forensic engineer to do an investigation to
determine if an electric cooking stove or an electric wall heater where
the deceased was found sitting against was the cause of a possible electrocution.
Upon examination of the scene by the writer, it was noted that the
room and all objects had been black-dusted for fingerprints. The body
had been removed as well as the television set, bed covering, clothes and
all personal effects.
Visual inspection revealed an older style golden colored fourelement cooking stove (with oven) unplugged and moved away from the
wall between the bed and kitchen counter wall. The stove had previously
been up against the wall and within approximately 1 ½ inches from the
125
edge of a built-in electric radiant wall heater. Examination of the stove
found that all five switches were in the “off” position. Dust and cobwebs
were found all over the rear of the stove suggesting no previous recent
tampering with the stovetop. The stove was grounded and connected to
a three-prong plug without a ground contrary to the manufacturers
instructions. While this is potentially hazardous, it was not a factor in
this case. One wire from one heating element was found broken off.
The rest of the stove remained unremarkable.
The wall-mounted air heater was grounded via a grounding stud.
Both spiral resistance-heating elements functioned and were unremarkable. The thermostat control was intermittent and clicks were heard
when the control knob was pushed in. The rest of the heater remained
unremarkable. The circuit breaker supplying the stove and the heater
was also unremarkable.
The local medical examiner had previously hired a local electrician
to measure for voltages between the stove and the heater. The electrician
measured 36-70 Volts ac. The victim was in contact with the stove and
the heater. The medical examiner’s investigation upon autopsy revealed
that the victim had red eyes and had almost bitten through his tongue
about ½ inch from its tip. The head and shoulders were burnt. There
was rectal defecation at the time of death. The previous day the victim
expressed great discomfort in breathing in a somewhat horizontal
position in a dentist chair. The victim’s heart was 700 g versus 350-375
g normal weight. He had a “rotten” cold and enlarged lymph nodes.
Above all, the victim had been consuming “Ma Huang” an Oriental
herbal remedy that contains Ephedra for weight loss.1
It was determined by this forensic engineer by use of an electronic
equivalent (CSA-STD) of a human impedance model that the voltage
between the stove and the heater was non-lethal, non-shocking and was
due to stray capacitive leakage currents in the insignificant micro amp
range. There was no defect in the stove or heater that would produce
shocking or lethal currents to a human body. As well, no evidence of
willful tampering or suspicious non-accidental conditions was found.
The coroner’s office later revealed that the probable cause of death
was due to the effect of “Ma Huang” on previous medical conditions.
This was not an electrical homicide.
Eric Rudolph who had been on the FBI’s Ten Most Wanted List at
the time of this incident was recently captured on May 31, 2003, in
western North Carolina.
Reference:
1 U.S. Food & Drug Administration and National Council for
Agricultural Health Fraud.
Electrocution, Homicide, Accident
C26
Latent Failures of Type NM Wiring
After Ground Energization
Mark E. Goodson, PE*, 1007 Shady Oaks, Denton, TX 76205
After attending this presentation, attendees will be able to describe
the changes in NM leakage characteristics following destructive ground
energization that can lead to both immediate and latent fire scenarios.
humanity by demonstrating that catastrophic failure can result from
energized type NM cables well after the insult actually occurred.
Article 240 of the NEC generally provides for overcurrent protection of conductors. Overcurrent protection is usually provided by a
Molded Case Circuit Breaker (MCCB) meeting the requirements of
NEMA AB-1. The underlying criterion for overcurrent protection is that
the protective device will always operate before energy dissipated in the
wire by resistive heating will damage the wire. The inverse time-current
relationship by which MCCBs operate form the usual basis for this prophylactic scheme. While Article 240 does provide protection for the normally operating facility, it is of no use when energization of neutrals or
126
grounds occur. In particular, there is no protection during the scenarios
commonly referred to as “floating neutral” and “energized ground”
occur. In these scenarios, current flow is unbounded because there is not
the usual corresponding increase in hot lead current.
We describe here several fires which have occurred as a result of
both the floating neutral and energized ground scenario. Research is then
outlined in which type NM cables were intentionally damaged by
injecting onto them ground fault currents for varied lengths of time.
Temperature rises on the cables were measured by both conventional
thermocouple techniques and by thermography. As part of this testing,
we measured changes in leakage. By applying AC power across the hot
and neutral leads, and then measuring leakage current in vector form
(Real + Imaginary), we were able to plot changes in both capacitance
and resistive current flows. Instrumentation for our labwork made use
of the IEEE488 bus, and a Vitrek dielectric analyzer. This vector
analysis is shown to be superior to conventional megger and hipot
testing in terms of its ability to predict failures while not violating Kelvin
criterion. Field measurement techniques used by maintenance workers
during damage assessment never attain the precision offered by vector
analysis.
As a result of the type NM testing and from empirical data, we are
able to demonstrate that catastrophic failure and arcing ignition can
result from energized type NM cables well after the insult has occurred.
In one scenario, a fire broke out within 3 weeks of the inducing of
current onto the ground. We describe the analytical tools used to show
causation, as well as the evidentiary items that should be searched for
when examining a structure for this type of fault. The usual source of
energization in these fires results when a neutral is accidentally pulled,
or when a utility company (during maintenance procedures) injects
current onto the ground lead. The most sobering part of this research is
that damaged cables can bring about ignition well after the injury has
occurred, and well after any open and obvious defects have been
repaired.
Fire, Floating Neutral, Dielectric Breakdown
C27
Swimming Pool Electrocutions Revisited
Harold E. Franck, MSEE*, Advanced Engineering, 4713 MacCorkle
Avenue SE, Charleston, WV 25304; James A. Ruggieri, PE, General
Machine Corp, 10710 Timberidge Road, Fairfax Station, VA 20039;
Darren H. Franck, BSCE, Advanced Engineering, 4713 MacCorkle
Avenue SE, Charleston, WV 25304
The goal of this presentation is to attempt to verify reported experiments as outlined in a paper titled, “Point Source Electrocution in
Swimming Pools and Spas,” presented at the 2002 AAFS meeting. The
tests reported in the paper could not be duplicated and did not conform
to theoretical results.
humanity by demonstrating an understanding by which forensic engineers evaluate the nature of swimming pool electrocutions.
Since 1990, the U.S. Consumer Product Safety Commission
(CPSC) recorded sixty deaths and fifty electrical shock incidents in or
around swimming pools owing to defective or improperly installed electrical equipment. In most of these cases, GFCI devices were not
included, and the predominating offending electrical appliances were
120 Volt AC pool lighting fixtures.
In either the home or industrial environment, the presence of water
clearly serves to augment electrical shock and electrocution risk. The
risk is produced by the reduction in human contact resistance brought
about by improved contact coupling between the victim and the electrical source via the water. The National Institute for Occupational
Safety and Health (NIOSH) observes that the presence of moisture from
environmental conditions such as standing water, wet clothing, high
humidity, or perspiration, increases the possibility of a low voltage elec* Presenting Author
trocution.1 However, despite a large amount of literature and research
describing shock risk and the water environment, there is still much misconception among electrical engineers regarding the mechanics of
shock in damp or waterwet environments.
Typically, the water used in swimming pools, hot tubs, or spas are not
ionic fluids and are thus considered poor conductors. As such, it is not
expected to see a substantial current flow between two poles immersed in
pool water, as the water provides a relatively high resistance path.
Mathematically, the field produced by a live 120 VAC conductor
immersed in a swimming pool is a boundary value problem and is best
described though Poisson’s equation. The boundary value problem
solution describes the field conditions. This equation is derived from
Maxwell’s Divergence equation. Maxwell’s Divergence equation in
point form is derived by the application of Gauss’ Law to an infinitesimal volume and is stated as:
Substituting into equation (1) for the electric field intensity E and
electric potential V, we obtain Poisson’s equation
In free space, ρ = 0 so that equation (2) reduces to Laplace’s
equation. The nature of the flow of current through a material determines
whether the material is a dielectric, conductor, or semiconductor. In
liquids, both positive and negative charges are free to migrate.
Generally, the conductivity of a liquid is given by
The first term represents the contribution to the conductivity from
negatively charged particles moving opposite to the E field and the
second term represents the contribution from positively charged particles moving with the E field.
According to Kraus2 water has the following conductivities:
Swimming pool water has few electrolytes and is therefore at best an
insulator or a poor insulator. Measurements of current flow through pool
water should not indicate appeciable amounts. Ground fault circuit interrupters (GCFI) have a threshold current of five milliamperes. Currents
above this value are recognized to pose a danger to humans in that they
may produce a disruptive effect on the equivalent electric dipole of the
heart. This effect can produce death.
The field configuration produced by a live conductor in a
swimming pool is dependent on its boundaries. This type of problem
may be solved by the application of Poisson’s equation, graphically,
experimentally, or with an analog or digital computer. Experimental tests
were conducted in a 20-foot by 25-foot swimming pool standardized at
neutral pH, requisite chlorine level, and room temperature. A voltage
source was supplied by immersion of a two conductor energized
extension cord near the sutface at a location near the submersible pool
light fixture to simulate a failed and hazardous ixture. A 5-foot by 5-foot
rectangular coordinate grid was formed. The grid provides voltages and
current measurement distance targets to help explain a field map of the
energy distribution in the x-y plane of the pool. A human model was simulated in accordance with IEC 479 and UL data to approximate a
nominal human surface area. The model, constructed from a twenty-four
guage galvanized steel sheet, buoyancy foam, insulators and resistors,
was positioned at the various grid nodes while voltage and current
measurements were recorded. The findings of the test show that for a
substantive shock risk and injury to occur to an immersed human
subject, certain physical contact must be made with an energized conductor, regardless of conductor immersion, conductor surface area, or
water chemistry, and the water container must present a definite ground.
The findings of this test could not reproduce the conclusions reached in
a previous AAFS paper and show substantive flaws in the reasoning and
the conclusions reached in that paper.
The voltage measurements along the pool grid were essentially
insignificant until the probe was very near the source. Similarly, current
measurements were in the microampere range under these conditions.
Essentially in this case, the tests indicate that physical contact with the
source and grounding of the individual is necessary to produce an electrical shock incident. The electric field produced in a water environment
is dependent on the energized surface area, the water chemistry, and the
characteristics of the ground.
References:
1. U.S. Department of Health and Human Services, Public Haelth
Service, Centers for Disease and Prevention, National Institute for
Occupational Safety and Health (NIOSH) Worker Deaths By
Electrocution - A Summary of NIOSH Surveillance and Investigative
Fingings, p.7.
2. Krauss, John D., Electromagnetics, Third Edition, McGraw-Hill,
p 123.
Electric Field Intensity, Conductor, Poisson’s Equation
C28
The conductivity of a liquid electrolyte is represented in the
diagram below and explains the movement of charges and the relative
direction of the fields with respect to the movements of the charges.
Safety Code Violations, Corrections,
and Replacements
Thomas P. Shefchick, BSEE*, Thomas P. Shefchick, PE, PO Box
62284, Sunnyvale, CA
The goals of this research project are to illustrate electrical code
violations, to demonstrate that code violations do not cause immediate
damage and to discuss repair or replacement of violating equipment.
This presentation will impact the forensic science community
and/or humanity by educating them about electrical hazards, safety,
damage and repair.
A video presentation of an electrical fire scene will be utilized to
illustrate electrical safety code violations. The damage caused by the
violations will be shown and the repair of the electrical system will be
discussed.
* Presenting Author
127
One evening the resident of a very fine home in a very exclusive
neighborhood returned home. The electric gate at the entrance to the
property worked okay. The driveway and property was lit as usual in the
evening. However, the resident noticed an unusual glow in the front
windows of the house’s piano room. Upon entering the house, smoke
was discovered but there was no alarm, since the house did not have
smoke detectors. The fire department was called to extinguish the fire.
Upon arrival the fire department found the fire emanating from electrical
wiring within the walls of the building. Normally, the fire department
turns off electrical power to a burning building before attacking the fire
because it presents a shock and electrocution hazard for the firemen.
The contaminants in water are good conductors of electricity and fire
hydrants are good ground conductors. This can result in serious injury
to firemen. However, the firemen had difficulty turning off the electrical
power in this burning building. After turning off the building’s main
circuit breaker, their thermal imaging equipment indicated that wiring in
the walls and beneath the building was still overheating and energized.
Subsequently, the fire department turned off the electric power at the
building’s remote electric meter box and extinguished the fire. The fire
department informed the residents of the fire building that the entire
electrical system of their home would probably have to be replaced.
The remote electric meter box was surrounded by poison oak. It
was corroded with vegetation and insects were found inside. Its cover
was not closed and its neutral was not grounded as required by the
National Electrical Code. Conductors were found attached to the two
pole 200 ampere circuit breaker in the distribution section of the meter
box, which fed into the meter box. These conductors were rated at 40
amperes and they would not be properly protected by a 200 ampere
circuit breaker. In addition, utility companies do not permit customers
to pull conductors through a meter box. The conductors went to a distribution box mounted on the left side of the meter box. However, the
conductors could have been used to bypass the meter and steal electricity. These conductors should have been noticed by the utility meter
reader and eliminated prior to the fire. Whoever installed the conductors
violated the National Electrical Safety Code by working in close proximity to energized electrical conductors. The illegal box attached to the
left side of the meter box was corroded with vegetation and had insects
inside. It fed electrical power to a duplex receptacle mounted on the
support for the meter box. The duplex receptacle previously supplied
power to outside wiring. Evidence of burning was found on the conduit
to the receptacle. Apparently, the outside wiring powered by the receptacle had overheated and caused a previous fire. Afterwards, the outside
wiring was powered by splicing into the main service cable to the
building in an underground splice box. The splice box was not sealed to
keep the elements away from the splices or the conductors, which were
not rated for outdoor or underground use. A number 12 AWG copper
wire, which is rated at 20 amperes by the National Electrical Code, was
found spliced directly to an aluminum cable rated at 200 amperes. The
200 ampere circuit breaker in the meter box would not prevent the 20
ampere wiring from overheating when current above 20 amperes passed
through it. The 200 ampere circuit breaker in the meter box was the only
overcurrent protection in the circuit. This explains why the house circuit
breakers did not de-energize the outdoor wiring. In addition, it is a violation of the National Electrical Code to splice copper conductors
directly to aluminum conductors. The dissimilar metals react and cause
connection problems. The outdoor splices were made approximately
four years before they failed and caused a problem. A neutral conductor
from the house panelboard had been brought out to the splice hole.
When the spliced conductors failed, they energized the neutral conductor
into the house. The energized neutral conductor in the house eventually
found a path to ground in a receptacle in the house. The amount of
current in the conductor/wire was only limited by the resistance of the
wire and its connections. Therefore, the current exceeded the rating of
the wire. The overcurrent caused the wire to overheat and ignite combustible materials located near it. Heat damage was also found in the
128
fire building’s main panelboard where the overheated neutral conductor/wire was located. This panelboard contained many electrical
code violations. It was not labeled, properly grounded or wired. A subpanel in the building was also defective. Wallcovering material had been
sprayed into it while its cover was off. The material would affect the
connections in the panelboard and the response of the circuit breakers to
overcurrent. Bad connections will overheat and result in electrical
breakdown and failure. Non-responsive circuit breakers will permit
overloaded electrical circuits to overheat and cause fires.
The repairs to the electrical system involved eliminating the code
violations. The wiring through the meter box was removed. The distribution section of the meter box was cleaned out and properly grounded.
The outdoor splice box was replaced with a sealed box rated for outdoor
use. Outdoor wiring was replaced with wiring rated for outdoor use.
The main panelboard in the house was replaced, correctly wired and
grounded. The subpanel in the house was replaced.
Code Violations, Electrical Damage, Electrical Repair
C29
Some Comments on the
Distinctions Between Experts
David G. Howitt, PhD*, University of California at Davis, Department
of Chemical Engineering and Materials Science, Davis, CA 95616
The goals of this research project are to provide some comment on
the alternative roles of an expert witness.
humanity by providing information about the role of the expert witness.
Anyone who has been asked the question as to the proportion of
work they do for plaintiffs compared to defense attorneys has probably
given some thought to this problem of “Seeing Both Sides of the Coin”
and wondered if it truly makes any difference. The ways in which I have
personally found them to differ are in the nature of the assignment and
the level of resources that are brought to the case. Typically as a plaintiffs expert one is trying to find out exactly what happened whereas as
an expert for the defense it is usually a case of simply defending against
the hypothesis. This is not always true because some defense attorneys
like to present an alternative scenario to a jury but in general the
approaches are rather different, the expert for the plaintiff is a detective
and the expert for the defense is a critic. Doing both is certainly more
interesting than being confined to one, but the task of the plaintiffs
expert is invariably more difficult and in most cases more rewarding
intellectually. The advantage to being a defense expert comes from the
fact that it is much easier to be critical than creative which brings us to
the subject of resources since despite this advantage there is invariably
more time invested by experts in the defense of a case than by the
plaintiff. Indeed things are usually on a much less grandiose scale when
working for the plaintiff and this probably has more to do with the distinctions in the way subrogation firms and defense attorneys conduct
business than legal strategy. The team concept is generally more
common as a defense strategy but is used by both, particularly when
multiple defendants or plaintiffs are involved and they pool their
resources. Even with large insurance companies as a plaintiff there is a
tendency to stress frugality presumably because the good name of the
organization is not at stake. Also when there are multiple experts they
tend to be brought in at different times in plaintiff cases but all at once
in the defense strategy, which creates a different type of working environment. Indeed I think it is fair to conclude that the work of the
plaintiff’s expert is not only inherently more difficult but it is also performed under more difficult circumstances.
A final question that one might address is whether there is any difference in the reliability of experts hired by plaintiffs as opposed to
defendants. We have all probably come across some pretty strange theories in the course of civil litigation, which raises some serious questions
about the professional competency of some experts, but the ones I have
* Presenting Author
come across have been about equally divided between the two camps.
Some examples that I will be happy to share include the spontaneous
separation of gases and the explosive defoliation of chickens but there
are also some examples where the scientific distinctions are so nice that
one could not confidently expect to be able to satisfactorily explain them
to a jury although they are nonetheless still very wrong. From my own
experience. However, I have generally found that about half of the cases
brought to me by plaintiffs have merit and this proportion is unchanged
in the cases that are also brought to me by defense attorneys. How this
reflects on the difference between being an expert for the Plaintiff and
opposed to the Defense is probably worthy of some discussion.
Expert, Plaintiff, Defense
C30
Arson/Murder - Expert Witnesses Biased Opinions
Marshall L. Smyth, BS*, Smyth Consulting Engineering, 809 N
Shepherd Hills, Tucson, AZ 85710
modification of current sex offender program protocols for correctional
populations to meet the growing number of sex offenders being identified in psychiatric centers. Enhanced program development centers can
better serve this population by providing an effective treatment protocol;
better serve the courts by proving more informed opinions regarding the
risk of re-offending; and better serve the community by safeguarding the
inappropriate discharge of individuals who are likely to re-offend.
humanity by demonstrating the severe consequences of prosecution fire
investigator overreach in determining the causes of two high profile
Arson/Murder cases in which the author testified as a defense expert
witness.
These two Arson/Murder cases involved different defendants (John
Henry Knapp in Arizona and Ernest Ray Willis in Texas), and widely
separated times. Each residence involved in the fire experienced
Flashover and Post Flashover burning in rooms of the residence. Nearly
all of the burnable materials in the rooms of origin were destroyed
during the fires.
The evidence and representative fire tests performed by the defense
after the initial trials and convictions indicate the prosecution’s fire
investigators misinterpreted the burn patterns and misidentified the
cause of each fire.
These experiences indicate that the misinterpretation of floor and
low level burn patterns resulted from one or a combination of the following:
1. Excessive freedom allowed experts in their testimony, now being
curtailed by the effects of:
a. Federal Rule of Evidence 702, Daubert v. Merrill-Dow
Pharmaceuticals case, Benfield v. Michigan Millers Mutual
Insurance Company case, and perhaps others
b. The advent of the National Fire Protection Association’s
“NFPA 921 Guide for Fire and Explosion Investigations” a
science based, consensus derived, authoritative guide, first
issued in 1992 and now undergoing its fourth update
2. Failure to perform representative corroborative tests or analyses
3. Failure to appreciate the dynamics of Flashover and Post
Flashover combustion
4. A lack of understanding of the impact of burning floor carpet,
carpet pad, combustible wall paneling, celotex ceiling tile and
furniture upholstery on the development of a room fire
5. Lack of engineering/scientific training and experience
6. Arrogance
7. The halo effect of being a member of the “Good Guys” team
ordained to rid the world of the “Bad Guys”
8. The pressure to convict applied by prosecutors
* Presenting Author
Two individuals were wrongly convicted of arson and murder.
Both were sentenced to death. Both have survived by the skins of their
teeth because of outstanding and very expensive pro bono efforts, in the
John Henry Knapp case by Attorneys Larry A. Hammond and Colin F.
Campbell of the law firm of Meyer, Hendricks, Victor, Osborn &
Maledon, Phoenix, Arizona, and in the Ernest Ray Willis case by James
S. Blank of the law firm of Latham & Watkins, New York, New York.
John Knapp was finally freed after three trials, many appeals, and a
post conviction evidentiary hearing.
Ernest Willis is still on death row after one trial, many appeals and
a post conviction evidentiary hearing. His case is being reviewed by the
Federal Western District Court of Texas, Pecos Division.
Fire Investigation, Expert Testimony, Biased Opinions
C31
Effects of Analytical Variability for
Calibrated vs. Uncalibrated PAH on
Hydrocarbon Source Recognition Methods
Jeffrey W. Short, MS*, Auke Bay Laboratory, National Marine
Fisheries Service, NOAA, 11305 Glacier Highway, Juneau,
AK 99801-8626
After attending this presentation, attendees will learn of implicit
limitations of hydrocarbon analytical data used for identifying hydrocarbon source contributions when applied to determining natural
resource damage assessment following accidental release of petroleum
hydrocarbons.
humanity by demonstrating that the limitations inherent in the usual
GCMS method used for analyzing environmental samples for evidence
of contamination by petroleum products are rarely recognized by experts
who interpret these data. The analytical reliability of the alkyl-substituted polycyclic aromatic hydrocarbons (PAH), which contain most of
the information for identifying hydrocarbon sources within the PAH
class of hydrocarbons, is substantially lower than for the parent homologues, but these data are usually interpreted on the implicit assumption
of equal reliability. This presentation will demonstrate how this lower
reliability constrains valid interpretations of these data, which should
help practitioners to avoid invalid conclusions regarding hydrocarbon
source contributions in natural resource or property damage assessments
from chronic or catastrophic releases of petroleum products.
Hydrocarbon source identification for samples contaminated by
petroleum products usually relies on data produced by gas-chromatograph/mass spectrometer (GCMS) systems. A single GCMS analysis
produces a wealth of data, and may include results for dozens of hydrocarbon analytes in each of three classes: the polycyclic aromatic hydrocarbons (PAH); the normal and branched alkanes; and the alicyclic
hydrocarbons, or biomarkers. Analysis of the biomarkers is often definitive, because these compounds are environmentally persistent and are
characteristic of the geologic age and conditions during formation of the
petroleum products in question. However, the concentrations of biomarkers in crude oil is low compared with the PAH or other alkanes, and
they may be absent entirely from the more volatile fractions of refined
petroleum products. This limits the utility of the biomarkers for hydrocarbon source resolution to environmental samples that are heavily polluted by crude oil or its less volatile refined products, such as the bunker
oils. In other cases, useful information for source identification may be
limited to the PAH and the other alkanes, and these may be altered by
weathering processes following introduction into the environment.
Weathering processes often lead to rapid degradation of the
alkanes, especially the normal alkanes, while alteration of PAH concentrations is usually slower and more predictable. Hence the PAH are
often the primary focus of efforts to deduce hydrocarbon source contributions in samples that are not heavily contaminated. Analytically, the
129
quality of PAH results may distinguished according to the availability
and use of authentic calibration standards. These standards are available
for many unsubstituted and for some alkyl-substituted PAH, permitting
fairly rigorous identification and measurement of these PAH in environmental samples. Unfortunately, the most abundant and persistent PAH
in crude and refined oils are the more substituted PAH, which comprise
multiple isomers for which authentic calibration standards are rarely
available. The identification, measurement and detection limits of these
alkyl-substituted PAH is therefore less certain, and their measurement
usually relies on response factors that are assumed to be identical to
either the un-substituted parent PAH or the most similar alkyl-substituted PAH for which a calibration standard is available.
Actual
detection limits for these uncalibrated alkyl-substituted PAH are necessarily higher than those of corresponding un-substituted homologues,
because the analytical signal is spread among numbers of isomers that
increase sharply with increasing alkyl substitution. The limitations
imposed by these considerations on efforts to deduce hydrocarbon
source contributions to contaminated environmental samples are largely
unknown.
In this presentation I compare the variability of calibrated and
uncalibrated PAH in a large series of reference samples for the Exxon
Valdez oil spill, to evaluate how differences in data quality between calibrated and un-calibrated PAH affect methods for assessing hydrocarbon
source contributions. The reference samples were all analyzed by the
same method at the same laboratory. Coefficients of variation for the
calibrated PAH are typically 10% - 15% for the calibrated PAH,
increasing to as much as 50% for the most substituted PAH. This
increasing variability constrains the resolution of both analyte ratio
methods and of multivariate statistical methods for deducing hydrocarbon source contributions. These limitations are illustrated by comparison with results for hydrocarbons in benthic and riparian sediments
from the northeast Gulf of Alaska (GOA). The hydrocarbons in these
GOA sediments may be from a collection of closely related sources that
preclude weathering alterations following introduction to the environment, and so provide a rigorous test of hydrocarbon source discrimination methods. Analyte ratio results from the reference sample set are
compared directly with those from the GOA sediments. Comparison of
results for a multivariate statistical approach, which makes much fuller
use of the available information than do analyte ratio methods, is accomplished by combining the Aitchison metric applied to hydrocarbon concentration pattern differences among samples, with hierarchical cluster
analysis. Results of these comparisons provide an example of the inadequacy of these approaches (especially the ratio methods) to detect
hydrocarbon source contribution differences that actually exist in the
field, when these sources are closely related geologically and the hydrocarbon analytes are limited to the PAH.
Hydrocarbon Source Identification, Polycyclic Aromatic
Hydrocarbon, Oil Spill
C32
Principal Component Analysis:
Mathematics and Voodoo
Willem A. Schreuder, PhD*, Principia Mathematica, 405 Urban Street,
Suite 305, Lakewood, CO 80228
After attending this presentation, attendees will understand the mathematical principles behind principal component analysis (PCA), where it
is appropriate to use PCA and how to debunk misuses of PCA.
humanity by demonstrating the proper application of PCA and by
debunking abuse of PCA.
Principal Component Analysis (PCA) is one of a number of methods
that can be used to reduce the dimensionality of data. The purpose for
reducing the dimensionality by PCA is to determine vectors of maximum
variation in the data, so as to better distinguish between data.
130
PCA has been used in the environmental field to distinguish compounds of different origins. However, as with many statistical techniques,
the practitioner is often poorly familiar with the technique and relies on a
computer program to perform the analysis. The purpose of this paper is to
describe the mathematics behind the technique, what it is that the computer programs do, and how to interpret the results.
A case study will be used to illustrate the proper and improper use
of PCA.
Principal, Component, Analysis
C33
A Baseline Study of Measurement
Uncertainty for Peak Ratios of
Isoprenoids and PAHs
Todd R. Crawford, BA*, Center for Toxicology and Environmental
Health, 615 West Markham Street, Little Rock, AR 72201; Denise A.
Sheppard, BS, Trillium, Inc., 2014 Carol Drive, Wilmington, DE 19808
when using ratios to identify or differentiate between oils and fuel types
it is necessary to take into account the variability or experimental error
inherent in the methodology(ies) used. Measured and potential error
terms for the use of peak ratios for identification and differentiation of
fuels will be illustrated and discussed.
The members of the forensic community attending will be more
wary of accepting at face value, ratio comparisons for the purposes of
identifying and differentiating between contaminating hydrocarbon
fuels. We hope to inform participants that there are questions of variability and experimental error that need to be addressed when interpreting the data.
Several methods for the identification of petroleum products have
been proposed and adopted by professional organizations recently
[ASTM 5739-00, Nordtest Method NT CHEM 001]. Peer-reviewed
journals have published papers describing similar methods for characterizing and differentiating oil sources. These methods generally
compare the relative abundances of two or more compounds, which are
assumed to be distinctive and characteristic of the samples: hereinafter
this approach will be referred to as comparing “peak ratios.” Peak ratio
methods have some acceptance within the scientific community, and the
results of peak ratio analyses have been used successfully in litigation.
However, no study has been presented which quantifies the uncertainty
of the measurements, and there are no criteria to compare peak ratios
within any of these methods. The fundamental questions, “How much
variation will be found in the peak ratios for the same material?” and
“How much difference must exist between peak ratios to distinguish different materials?” have not been addressed.
This presentation will describe our investigation of the uncertainty
of peak ratio measurements for isoprenoids (nor-pristane, pristane,
phytane), and for polycyclic aromatic hydrocarbons (PAHs) and their
alkylated homologues. Isoprenoids, a group of branched alkanes structurally based on isoprene, do not biodegrade readily in the environment
and are therefore often used to distinguish middle distillate fuels (diesel,
heating oil, fuel oil). AHs are cyclic compounds based on conjoined
benzene rings. It is generally assumed that the larger PAHs (four or more
rings) are highly resistant to degradation in the environment. PAH patterns which can be compared as a series of ratios have been used to distinguish between samples of different oil sources.
Ratios are usually calculated based on peak heights or areas from a
gas chromatogram, generally using a flame ionization detector (GCFID) for the isoprenoids, and a mass spectrometer (GC-MS) for the
PAHs. All instrumental analyses are subject to experimental error.
Analyzing the same sample or sample extract multiple times on the same
instrument will produce a range of results. The range of results, and
hence the uncertainty of the peak ratio measurements, increases as other
variables are introduced to the analysis, e.g., sample extraction, sample
* Presenting Author
concentration, instrumental method, data analysis methods, etc. This
investigation evaluated the uncertainty of peak ratios with respect to
concentration, instrumental methods, and data analysis methods for GCFID and GC-MS analyses.
The GC-FID investigation evaluated the measurement uncertainty
for the peak ratios of the isoprenoids nor-pristane, pristane, and phytane
in a regular automotive diesel fuel (Texaco). The Texaco fuel was used
as a reference material in our laboratory for three years. It was analyzed
at concentrations ranging from 1000 to 50 µg/g (ppm) on two GC-FID
instruments.
The GC-MS investigation evaluated the measurement uncertainty
for the peak ratios of the PAHs, parent compounds and alkylated homologues in various crude oils. Multiple Agilent GC/MSD systems were
used to perform the analyses in two different laboratories.
Good Laboratory Practice (GLP) requires that results be reported
with an “error term” (the result obtained, plus or minus the statistically
determined measurement uncertainty for the analytical technique). Error
terms readily show the variability inherent in a measurement procedure.
When peak ratios are used to differentiate between materials, the reliability of the numbers used to generate the ratio must be assessed. One
ratio calculated from the heights of two peaks in a single chromatogram
of one sample at a specific location does not address variability: that
single ratio cannot reliably be compared to a similarly generated ratio for
another sample to distinguish different contaminants.
The Federal Rules of Evidence include, under Rule 702, assessment
of whether testimony is based on methodology that is scientifically valid
and can be properly applied to the issue at hand. Considerations bearing
on this assessment include not only peer review, publication, and existence of standards, but also the known or potential error rate for the
methodology used.
While peak ratios can be a very powerful tool for forensic environmental chemistry, in order to be scientifically supportable and defensible
an explicit statement of the measurement uncertainty must be included
with the result.
PAHs, Ratios, Isoprenoids
C34
Benzofluorene/Methylpyrene Ratios
as a Source Identification Tool
David R. Craig, BS* and David M. Mauro, MS, META Environmental,
Inc., 49 Clarendon Street, Watertown, MA 02472
The objective of this paper is to present data in support of the use
of benzofluorenes and methylpyrenes as a means of tracing environmental contamination to its source.
humanity by describing how to improve the accuracy and reproducibility
of environmental source identification.
Polycyclic aromatic hydrocarbons (PAHs) often drive costly site
investigation and remediation work. Former manufactured gas plant
(MGP) sites frequently are contaminated by PAHs from one or more
sources depending on their operational histories, and in many cases the
nature of environmental work at MGP sites can depend on identifying
the sources of the PAHs. However, identifying the sources of PAH contamination at MGP sites is complicated by the numerous possible PAH
origins. Common anthropogenic sources of PAHs include coal tar and
coal tar products, refined petroleum products, MGP wastes, exhaust
from heating systems, vehicular emissions, and others. PAHs also are
created naturally by forest fires and, in some cases, synthetically by bacteria and algae. As PAHs weather and commingle with PAHs of other
origins, as is typically the case in industrial soils and urban sediments,
identifying their sources proves increasingly difficult.
Current approaches for identifying sources of PAHs in soil, groundwater, sediments, and surface water at former MGP sites have largely
relied on the molecular fingerprint or the relative abundance of indi* Presenting Author
vidual PAHs or groups of PAHs1. However, this approach suffers from
two major problems: first, there are many sources of PAHs and their
PAH patterns can be very similar, and second, environmental weathering
(dissolution, evaporation, and chemical and biological transformations)
can alter the PAH patterns and confound their interpretation. Recent
studies have shown that certain ratios of PAHs and alkylated PAHs are
relatively insensitive to environmental weathering processes and can be
used to distinguish separate sources with similar PAH patterns even in
weathered samples. For example, the ratio of benzo(b+c)fluorene to total
monomethylpyrenes was found to vary little in coal tar-contaminated
sediments that had weathered to various degrees (unpublished). The benzofluorenes/methylpyrenes ratio (BF/MP) depends on PAH formation
conditions and is measurably different among different PAH sources.
For example, the BF/MP ratio of a coal tar sample was found to be
approximately 3.9 while that of a former MGP tar was about 1.5.
This presentation reports some of the results of research to identify
effective environmental forensic methods for application at former MGP
sites.1 The application of extended PAH profiles (EPA 8270 modified)
and the use of various PAH ratios, especially BF/MP ratios for the determination of PAH sources in non-aqueous phase liquids (NAPLs), soil,
and sediment will be discussed.
Reference:
1. “Chemical Source Attribution at Former MGP Sites,” EPRI Technical
Report 1000728, December 2000.
PAHs, Environmental Forensics, Source Identification
C35
When is a DNAPL a DNAPL?
PA 19320
USEPA rule of thumb for the presence or absence of a DNAPL is not
always valid.
humanity by demonstrating the need for an environmental forensic scientist to use analytical chemistry methodologies that detect and identify
hydrocarbons associated with chlorinated organic chemicals.
The USEPA and hydrogeologists have for a considerable length of
time considered the presence of a chlorinated solvent as a dense nonaqueous phase liquid (DNAPL) in an aquifer when the groundwater concentration of the chemical exceeds 1% of the equilibrium solubility of
that chemical.1 For example, the equilibrium solubility of
trichloroethene (TCE) in water is 1,100 mg/L (ppm). Therefore, if TCE
is found above a concentration of 11,000 µg/L (ppb) in a monitoring
well, then it is reasonable to conclude that the TCE is present in the
aquifer as a DNAPL. Certainly, if the TCE is a DNAPL and is sinking
through the aquifer material, then remediation is going to be difficult at
best and impossible at worst.
But, is the high “hit” really a TCE DNAPL? Everyone seems to
agree that TCE is more dense than water with a density of 1.47 g/mL.
Also, most environmental professionals know that TCE and other chlorinated solvents have been and are used as degreasers. Many of these
degreasing uses have been in metal fabricating industries where the
chlorinated solvents were used in vapor degreasers. This use was
usually followed by a recycling of the solvent by distillation leaving a
hydrocarbon-solvent mixture waste. If this mixture is the material in the
environment, then there could be three completely different scenarios:
1. The mixture density is less than 1 g/mL. This material is a light
non-aqueous phase liquid (LNAPL). The LNAPL contains TCE, for
131
example, at concentrations exceeding 11,000 µg/L yet it is not a
DNAPL.
2. The mixture density is greater than 1 g/mL. This material is a
DNAPL, but it will act differently than a pure TCE DNAPL. For
instance, a TCE-alkane wax mixture will act like a DNAPL with a high
viscosity whereas pure TCE would move further and faster due to its low
viscosity.
3. The mixture starts being transported on the water table as an
LNAPL because the mixture has a density less than 1 g/mL. As the
mixture is transported, the hydrocarbon aerobically biodegrades and the
density of the mixture changes to a value greater than 1 g/mL and begins
to sink into the aquifer. The smear zone can promote this phenomenon.
The environmental professional should always test the hypothesis
that there may be a hydrocarbon portion of a chlorinated solvent release
to the environment. Remediation costs depend on it. Several case
studies will be discussed where high concentrations of chlorinated solvents in groundwater are not due to DNAPLs. Also, the use of the
chemical concept of partition coefficient will be used to show the fate
and transport of the chlorinated solvent.
Reference:
1. USEPA Evaluation of the Likelihood of DNAPL Presence at NPL
Sites National Results, EPA 540-R-93-073, PB93-963343, page xi,
September 1993.
Chlorinated Solvents, DNAPL, LNAPL
C36
Is Amorphous Silica Gel Non-Toxic?
Differences in Dose: Exposure Route
and Physical Form
James S. Smith, Jr., PhD*, Oak Creek, Inc., 60 Oak Creek, Buxton,
ME 04093-6616
After attending this presentation, attendees will have a better understanding of how the toxicity of a substance changes with route of
exposure and physical form. Dr. Smith will use two case studies
involving amorphous silica gel (“ASG”) or silica aerogel (“SA”) to illustrate how changes in the route of exposure and a material’s physical form
can alter its ability to cause adverse health effects.
humanity by demonstrating how information relating to exposure route
and an agents physical form can determine the type and severity of toxicological effects observed in exposed people. Understanding these concepts is essential for regulators involved in the evaluation of agents for
the protection of human and environmental health, interpretation of regulations concerning pesticide use, and in the use of expert witness testimony for effective risk communication. This presentation provides a
basis for understanding these concepts, applying them to the evaluation
of compound toxicity, and for use in risk communication.
This paper has three objectives: (1) to define the qualities of dose
that makes a substance a poison; (2) to illustrate how toxicity of ASG is
determined by route of exposure and physical form; and (3) to illustrate
how such dose information can be abused in legal proceedings.
Toxicologists often say that “the dose makes the poison.” This
implies that all substances are toxic and that it is the magnitude of the
dose that determines whether a substance is either safe or toxic.
Although true, there is much more information required about the “dose”
of a substance that determines whether a substance is a poison than
simply its magnitude.
In the last 30 years of cleaning up hazardous waste sites, federal
and state regulators and even some toxicologists have confused the
amount of a substance in environmental media with the dose. By definition, dose is the amount of a substance that reaches the organ (“target
organ”) where the material has the toxic effect of interest. Consequently,
132
dose is often adjusted to reflect the fraction of the substance in soil or
water that is able to enter the body (i.e., bioavailability) and reach the
target organ of interest. For example, generally less than 25 percent of
the arsenic found in soil is absorbed form the gastrointestinal tract after
ingestion. Furthermore, it is well recognized that the amount of a substance that is able to reach the target organ differs with different routes
of exposure (i.e., ingestion, dermal contact, inhalation, and injection).
For example, where nearly the entire injected dose of barium reaches the
target organ, less than 10 percent is absorbed through the skin. Finally,
the physical form of a substance can have as much to do with its relative
bioavailability and toxicity as does the magnitude of its dose. For
example: although the U.S. EPA assumes that all arsenic compounds
have toxicity equivalent to the mass of arsenic in those compounds, their
toxicity differs with respect to their relative solubility. In another
example, only asbestos fibers of a certain size are associated with the
development of cancer in the lung. This presentation discusses how the
route of exposure and the physical form of ASG can combine to result in
adverse health effects.
ASG is sold to end users as a non-toxic “natural” substance with a
variety of uses, including use as a food product additive, in paints, cosmetics and in pharmaceuticals, desiccant, and pesticide. For the vast
majority of these uses ASG is essentially without toxic effect to the end
user, but for at least one specific use, ASG poses a health hazard to
people. Although silica is a “natural” element in the environment that is
commonly associated with beach sand, soils, and rock, ASG is a manmade product that is radically different from the many silica products we
are familiar with (i.e., window glass, ceramics, and silica wafers used in
the electronic industry). Toxicologists have long recognized the seriousness of the health consequences associated with crystalline silica
exposure; silicosis. Because chronic exposure to ASG exposure does not
result in silicosis, many business concerns have labeled it “non-toxic.”
Dr. Smith explores ASG toxicity under different exposure routes
and physical forms commonly used in product formulations. Dr. Smith
uses two case histories to illustrate how dose information is abused in
legal proceedings and proposes several approaches to educating jurors to
minimize dose information abuse.
Amorphous Silica Gel, Silica Aerogel, Silica
C37
Air Samples Collected in Tedlar® Bags
Engrid S. Carpenter, BS*, Trillium, Inc., 9312 Highland Gardens
Road, Baton Rouge, LA 70811
implications of sample handling, storage conditions, and holding times
on samples collected in Tedlar bags and Summa canisters.
The impact of this presentation is to allow for a more informed
choice regarding the use of Tedlar® bags or Summa canisters to accurately and cost effectively assess a site.
The purpose of this paper is to present results for air samples collected in Tedlar® bags and Summa® canisters in support of an on-going
monitoring program. Implications of sample handling, storage conditions, and holding times on both sampling devices will be discussed.
A three-day holding time from the date of sample collection is specified for all samples collected in Tedlar® bags. Air samples collected in
Summa® canisters are considered stable for up to fourteen days from the
date of sample collection. Vapor permeation values are provided for
many of the volatile organic compounds collected in Tedlar® bags
because the manufacturers are aware that permeability is inherent to this
collection device. Summa® canisters are not permeable. So, why would
you use Tedlar® bags to collect air samples? Usually costs are the
deciding factor. Tedlar® bags can be purchased at a fraction of the cost
of Summa® canisters. In addition, shipping of Tedlar® bags is signifi-
* Presenting Author
cantly less than the cost of shipping the much heavier Summa® canisters. Tedlar® bags for trace analyses are disposed after use while
Summa® canisters must be decontaminated after each use and shipped
back to the field, both of which incur additional charges. But are we
sacrificing a true assessment of a site to save money?
For an on-going project, samples of untreated influent and treated
effluent are collected on a daily basis in Tedlar® bags or Summa® canisters. Method TO-15 is used to analyze the samples for chloroform and
carbon tetrachloride. Occasionally, Method TO-14A was used to analyze
the samples when the dedicated Method TO-15 instrument was not functioning properly. A reporting limit of 0.50 parts per billion by volume
(ppbv) was used for both target analytes and a calibration range of 0.50
ppbv to 40 ppbv was employed for this project. Many of the samples for
this project required significant dilution to obtain concentrations of
carbon tetrachloride within the established calibration range of the
instrument.
Initially, the daily influent and effluent samples were placed in the
same sample cooler for shipment to the laboratory. When collected in
Summa® canisters, concentrations in the influent sample were consistently and significantly higher than in the effluent sample demonstrating
that the facility was using an effective treatment process. However,
when the daily samples were collected in Tedlar® bags and shipped
together in the same sample cooler, the target analyte concentrations
were very similar in the influent and effluent samples. This suggested
that either the treatment process was not working properly or that an
equilibration of chloroform and carbon tetrachloride concentrations was
occurring between the Tedlar® bags. The client was requested to place
each Tedlar® bag in its manufacturer’s original box immediately after
sample collection. When the separate boxes were submitted to the laboratory for analysis, the concentrations of the target analytes in the
influent samples were again consistently higher than in the effluent
samples.
Subsequently, during the course of the project, it was noted that the
concentrations of chloroform and carbon tetrachloride were again very
similar in the influent and effluent samples collected in Tedlar® bags.
The client was contacted and we were assured that the samples collected
in Tedlar® bags were being submitted to the laboratory in separate
boxes. The laboratory was then contacted to see if any changes had been
made to the laboratory sample-handling process. It was discovered that
during sample log-in, the Tedlar® bags were currently being removed
from their boxes and the samples were placed side-by-side in the air lab
until analysis. The laboratory was requested to replace each Tedlar® bag
in the box it was received in until analysis. After this procedure was
implemented, the concentrations of the target analytes were again consistently higher in the influent samples.
Using four years of data, we have also observed declining concentrations of carbon tetrachloride between samples analyzed within one
day of sample collection and the same samples re-analyzed between two
and six days after sample collection. In some cases, the concentration of
chloroform increased, suggesting degradation of the carbon tetrachloride, but in most cases the concentrations of both carbon tetrachloride and chloroform decreased, suggesting loss of these volatile
compounds from the Tedlar® bags. No losses of similar magnitude were
noted in the samples collected in Summa® canisters. It is apparent that
Tedlar® bags are far more permeable than most people think
To increase the chances of getting an accurate measure of the target
analyte concentrations, samples collected in Tedlar® bags should be
shipped in their original boxes, scheduled for next day receipt at the laboratory, and analyses should be performed as soon as possible after
sample receipt. To minimize cross contamination as well as loss of
volatile organic compounds, we feel the use of Summa® canisters is far
preferable for the collection of air samples.
Tedlar® Bags, Summa Canisters, Air Samples
* Presenting Author
C38
Vapor Pathway Measurement
Strategies/Lessons Learned
Dominic L. DiSalvo, BA, ME*, Malcolm Pirnie, Inc., 104 Corporate
Park Drive, White Plains, NY 10602
After attending this presentation, attendees will have an understanding of sampling and analytical strategies used to measure volatile
organic compounds in soil vapor and crawl spaces below occupied
buildings.
humanity by demonstrating the importance of developing detailed sampling and analytical strategies for measuring vapor pathways.
This presentation will provide an overview of sampling and analytical strategies used to measure and evaluate volatile organic compounds (VOCs) in potential vapor pathways from contaminated groundwater/soil to indoor air. These strategies are based on experience gained
by Malcolm Pirnie in vapor pathway measurements on a wide array of
sites with groundwater and soil VOCs. Unfortunately, the same VOCs
(such as tetrachloroethylene – PCE) that are found in contaminated
groundwater and soils may also be found in the occupied space in
buildings from other commonly used sources, such as dry cleaning
brought home by a homeowner. This complicates the identification of
the source of the VOC in the indoor air and its impact on the occupants.
Therefore, indoor air sampling is considered a last resort, and is typically
preceded by extensive subsurface soil vapor investigations that can
assess a VOC levels below the ground surface.
The first step in developing the sampling and analytical strategy
was to establish the criteria to be used to evaluate if a vapor pathway
exists and if so, what information is needed to estimate indoor air quality
and potential health risk. Health risk-based VOC screening levels for
indoor air and attenuation factors for projecting the impact of soil vapor
on indoor air quality were determined, then the laboratory requirements
for vapor analyses, including methodology, method detection limit
(MDL), and quality assurance/quality control (QA/QC), could be established. Since the MDL selected for the project was below the parts per
billion range by volume, special consideration was given to the sampling
train (Summa canister with accoutrements) selection and laboratory certification process to assure cleanliness.
The second step was to understand the site and local area environmental conditions that can cause or contribute to a vapor pathway. These
included soil conditions down to groundwater, building construction,
subsurface preparation (backfill), utilities, proximity to the contaminated
groundwater or soil, and meteorological conditions. Once these environmental conditions were understood and screening levels established,
Malcolm Pirnie developed a sampling plan to collect representative
samples of air located below the building and in ambient air and, on
occasion, inside of buildings. This included sampling the air in open and
closed crawl spaces beneath structures plus soil vapor in soils beneath
slab-on-grade residential/industrial foundations using soil probes angled
beneath the foundations or inserted below the foundation through holes
drilled through the foundation. The sampling location selection and
preparation was crucial to establishing representative samples of
existing vapor pathways with minimum disturbance of on-site
conditions.
The implementation of the sampling and analytical plan required
significant logistical coordination and project oversight. The preparation of clean sampling trains and their monitoring during sampling
were critical to the success of the plan. Integrated sampling (over 8 to
24 hours) limited potential room for error in sampling train flow calibration and tightness testing. Outside sampling also was influenced by
weather conditions. The cases that will be discussed allowed Malcolm
Pirnie to test the sampling trains to their extremes while providing the
needed analytical data.
Malcolm Pirnie has used the expertise gained sampling vapor
pathways on contaminated sites to develop more efficient, effective and
133
representative sampling and analytical strategies. The lessons learned
from vapor pathway measurements have been shared with regulatory
agencies and should assist the profession to provide guidance on
sampling and analysis.
Vapor Pathway Measurements, Soil Vapor, Gas Screening
C39
When Timing Is Everything:
How Historical Aerial Photography
Established Timing of Industrial Activities
Resulting in Environmental Damage
Kristen K. Stout, BA*, Environmental Research, Inc., 5267 John
Marshall Highway, Suite C, Linden, VA 22642
value of using historical aerial photography in environmental litigation.
The presentation will introduce the community to a cost-effective,
unbiased source of information that can be used to determine the timing
of environmental damage for allocating costs associated with environmental remediation.
Learning Objectives: This abstract presents two examples in
which historical aerial photography was used to determine the timing of
environmental damage in Areas-of-Concern (AOCs) at a large chemical
manufacturing facility.
Method: Comparative stereoscopic analysis of a series ofhistorical
aerial photographs to determine when specific chemical manufacturing
activities started. Using geo-referenced digital images to precisely track
changes in the deposition and removal of materials over time to
determine when contaminated soil was placed in a specific area.
This abstract illustrates how the timing of waste disposal and industrial activity at a major chemical manufacturing facility was determined
through detailed stereoscopic analysis of historical aerial photography.
The photographic analysis was conducted in support of litigation
resulting from the chemical company’s claims against insurance companies for costs associated with environmental investigations and remediation. The insurance policies were in effect during specific time
periods; claims associated with activities or events outside those time
periods would not be covered.
The chemical manufacturing facility is located in the Mississippi
River flood plain in Louisiana. A series of levees, canals and ditches
control periodic flooding that occurs along the river. Operations at the
facility began in 1958 with chemical production and storage infrastructure located in the southern and southeastern portion of the
property. Over the course of several decades, additional production
plants and storage infrastructure, rail lines, and canals were constructed,
primarily moving in a north to northwesterly direction across the
property.
Sixty-five dates of photographic coverage spanning the time period
from 1952 to 2001 were acquired and reviewed to document features and
conditions of interest and to study change over time. Many of the photographic images were geo-referenced to aid in a detailed sequential
comparison of specific areas.
The aerial photographic analysis focused on specific AOCs at the
facility. One of the major AOCs was a vinyl chloride plant reportedly
constructed in the mid- to late 1970s. Leaks in storage and feed tanks
were discovered in the late 1980s and early 1990s, along with free phase
product in the ground. The company asserted that the leaks resulted
from faulty tank construction and would have occurred shortly after the
plant went on-line; however, it did not provide any definitive documentation as to when the plant went on-line.
Based on the analysis of several dates of aerial photography flown
in the 1974 to 1980 time period, it was concluded that the plant was still
under construction as late as July 1977. High resolution aerial imagery
flow on July 6, 1977, provided the critical visual evidence. Various plant
134
structures and pipe galleries were not completed. Stacks of pipes, beams
and other construction material were visible in many locations around
the new vinyl chloride plant, along with construction equipment.
Furthermore, two impoundments at the plant were still under
construction.
As of July 6, 1977, much of the ground surrounding the structures
appeared rough and uneven, with several mounds of earthen material. In
later dates of photography, the plant yard had been graded and a cover
material of some type (probably gravel or crushed shell) had been applied.
The insurance policy ended before July 6, 1977 (i.e., before the
plant went on-line); and therefore, the release occurred outside the
policy period.
Another AOC was located south of the main chemical manufacturing facility on the river side of a levee. Contaminated soil was discovered adjacent to two borrow pits. The two borrow pits were separated
by a narrow strip of land that served as an access road from time to time.
The entire area was flooded on a regular basis and was highly susceptible to erosion during these periods.
In 1986, the company discovered contaminated soils in the area
between and to the east of the two borrow pits. A portion of the area was
excavated and backfilled. Analysis of the contaminated soil indicated the
presence of organic compounds. Three follow-on soil boring investigations were conducted in 1986, 1987 and 1993 to define the vertical and
horizontal extent of the contaminated soil.
The company indicated that sporadic dumping had occurred in the
general area located along the riverside of the levee during the 1950s and
1960s. Initial review of historical aerial photographs from the 1960s
showed filling and dumping of unconsolidated material in several locations. However, further analysis of later dates of photography revealed
that this fill material was physically removed and/or eroded away during
river flooding.
The approximate areal extent of the contaminated soil, as derived
from maps produced during the three investigations, was registered to a
1984 geo-referenced digital image of the AOC. This was the closest date
of aerial photography that pre-dated the initial investigation and subsequent partial source removal. The resulting composite map was digitized
to create a geo-referenced spatial foot print of the contaminated soil.
This spatial foot print was overlain to earlier dates of geo-referenced
imagery, from which changes in the terrain due to anthropogenic activity
and natural forces were tracked. From this detailed and precise comparison of sequential dates of aerial photography, it was ascertained that
the contaminated soil identified in the investigations was placed in the
AOC in the early 1980s, and possibly, to a lesser extent, in 1974. This
was significantly later than originally purported.
These are but two examples in this case where detailed analysis of
historical aerial photography was used to determine the timing of environmental damage for environmental cost recovery litigation.
Aerial Photography, Environmental Damage,
Cost Recovery Litigation
C40
Passing the Buck (Around)–Using
Forensic Environmental Evidence to
Allocate Environmental Cleanup
Responsibility and Costs Among
Potentially Responsible Parties
John B. Robertson, BS*, John B. Robertson Consulting, 40107 North
3rd Street, Desert Hills, AZ 85086
The goal of this presentation is to demonstrate concepts and
parameters for applying technical environmental investigation
approaches and results to establishing the relative degree of responsibility among parties responsible for environmental harm at Superfundtype sites, using right and wrong examples and case histories.
* Presenting Author
humanity by providing help to environmental site investigators and
those involved with developing and negotiating cost allocations for
Superfund-type sites with constructing and applying the most appropriate, justifiable, and equitable technical basis for allocation and to
acquaint listeners with some possible new and innovative approaches.
Allocation of responsibility and response cost among potentially
responsible parties at federal Superfund and equivalent state-program
environmental cleanup sites is probably the most contentious and litigious aspect of those programs. The process is made difficult and
complex for several reasons, including: parties rarely are willing to
accept any responsibility for an abandoned contaminated site (partly
because of the joint and several liability aspect of Superfund); every
party that does accept some responsibility naturally wants their share to
be the smallest; CERCLA and state regulations provides little guidance
on how allocations should be done; every site is unique in the circumstances that led to the state of contamination and the history of owners,
operators, generators and transports; every site is different in terms of
the technical evidence that reveals the history and causes of contamination driving the cleanup.
Despite these difficulties, responsible parties can usually be made
more accepting of responsibility and of a fair allocation share if an
appropriate allocation model is developed, based on equitable principles
and defensible technical parameters, that can be reasonably supported
with site-specific environmental evidence. This presentation will explore
some successful and unsuccessful allocation models and why they succeeded or failed, together with the types of forensic evidence that has
proven to be most useful in allocation cases.
The most fundamental principle driving fair and successful allocations is that of cost causation: the parties that contributed most to the
environmental harm that is driving cleanup costs are the parties that
should pay the most. Often, a simple volumetric allocation is attempted,
if appropriate evidence exists. However, seldom is sufficient waste
volume evidence available to allow a volume-only based allocation to be
done fairly. Other parameters than can be useful in the allocation
formula include time of involvement in the site, relative portion of the
site occupied or used, types of activities at the site, waste management
practices, timing of waste releases at the site, and characteristics of the
hazardous materials/wastes handled at the site.
The greatest challenge, of course, is developing convincing evidence that indicates each party’s relative role in causing the harm. That
is where the bag of forensic investigation/analysis tools comes into play.
Some of the investigative tools and types of evidence that have proven
to be most useful in developing and applying allocation models are the
following:
• determining three-dimensional distribution of key contaminants
in soil and groundwater
• reconstruction of the historical spread of contamination from
sources using historical data and computer models
• using historical aerial photos to identify source areas, parties,
times of releases, and other evidence
• interviewing former employees and other witnesses to past
activities
• using information from similar analogue sites
• researching old operational manuals, policies, and guidance
documents
• applying geographic information system (GIS) technology to data
compilation, analysis, and display
• assessing relative toxicity, mobility, and persistence of different
contaminants in the mix
Several interesting case histories will be used to demonstrate how
these and other lines of evidence have been used and misused to allocate
cleanup responsibility and costs.
C41
Cosolvency
PA 19320
water insoluble organic chemicals are rarely made more soluble in water
when co-deposited with a water-soluble solvent(s).
humanity by explaining that higher than expected concentrations of a
water-insoluble organic chemical in water should not be attributed to
cosolvency without extraordinary proof.
Cosolvency has been used to explain why water-insoluble organic
compounds are found at concentrations significantly above their equilibrium water-soluble concentrations in groundwater. This has been
especially true for polychlorinated biphenyls (PCBs). For example,
assume that PCBs and a chlorinated solvent such as trichloroethene
(TCE) were discarded together. Now, in a groundwater sample from the
monitoring well, one finds concentrations of PCBs that exceed the water
solubility for PCBs. The explanation used for the higher than expected
concentrations of PCBs is cosolvency. In other words, it was assumed
that higher concentrations of PCBs such as Aroclor 1260 in water were
caused by the ability of TCE or another solvent to carry the PCBs into
the water solution and hold the PCBs in a soluble form.
However, research and recent publications1,2,3,4 show that cosolvency rarely occurs and certainly not with TCE and PCBs. What does
occur is that the solvent “salts out” or decreases the water solubility of
the PCBs. The equilibrium solubility of PCBs is lowered by the presence
of a solvent in the groundwater until the solvent reaches percent values.
The solvent does not increase the solubility of insoluble materials such
as PCBs in the groundwater.
Is that the end of the story? Of course not. In another example, the
Stringfellow lagoons above Glen Avon, California, contained cutting oils
that migrated through the aquifer. How is that possible? Cosolvency is
the answer here. At the Stringfellow site, there were high concentrations
of detergents from water-soluble cutting oils which were used to solubilize oil in water. They continued to do their job in the aquifer material.
Although these oils were never reported on the laboratory data sheets
from analyses of the water being transported through aquifer material
from the lagoons toward Glen Avon, they were confirmed to be present
based on the total ion chromatograms from the semivolatile organic
compounds analyses.
It is obvious that forensic investigations need to consider cosolvency, but the regulatory analytical chemistry tests required for an investigation are not always the tests necessary to demonstrate cosolvency.
Cosolvency must be carefully proven scientifically before it is used as an
explanation for the forensic investigation.
1 Coyle, G.T., T.C. Harmon, and I.H. Suffet, “Aqueous Solubility
Depression for Hydrophobic Organic Chemicals in the Presence of
Partially Miscible Organic Solvents,” Environmental Science &
Technology, Vol. 31, pp. 384-389, 1997.
2
Munz, Christoph and Paul V. Roberts, “Effects of Solute
Concentration and Cosolvents on the Aqueous Activity Coefficient of
Halogenated Hydrocarbons,” Environmental Science & Technology,
Vol. 20, pp. 830-836, 1986.
3 Groves, Jr., Frank R, “Effect of Cosolvents on the Solubility of
Hydrocarbons in Water,” Environmental Science & Technology, Vol. 22,
pp. 282-286, 1988.
4 Ulrich, Glenn, “Fate and Transport of Ethanol-Blended Gasoline in the
Environment,” Governer’s Ethanol Coalition, pp. 15-16, October 1999.
Cosolvency, Water Solubility, Salting Out
Environmental Cleanup, Cleanup Cost Allocation,
Environmental Forensics
* Presenting Author
135
C42
An Adaptive Sampler for Human Scent
Measurements in Field Forensics
Brian A. Eckenrode, PhD* and Valerie J. Cavett, BA, Federal Bureau
of Investigation, FBI Academy, Building 12, Quantico, VA 22135; Scott
A. Ramsey, BS, Michigan State University, School of Criminal Justice,
East Lansing, MI 48824
After attending this presentation, attendees will understand several
approaches to volatile organic compound analysis in the field for
forensic purposes, as well as new instrumentation developments that
facilitate these analyses.
humanity by demonstrating how the miniaturization of detectors will
help drive the commercialization of small, low power, light-weight
instruments for near real-time field analyses.
The FBI’s laboratory division and other operational units within the
FBI require rapid and reliable volatile organic compound (VOC)
analyses in the field to assess human scent collection instrumentation
and improve their overall response strategy and effectiveness for
chemical vapor determinations. Trace detection of VOCs captured via
scent pads or emanating from clandestine human burials in the field has
been difficult and current methods rely heavily on the use of canines.
Because canines are subject to exhaustion and maintenance an improved
methodology for screening large areas is required.
The Counterterrorism Forensic Science Research Unit (CTFSRU)
has developed a new adaptive sampling and analysis approach to trace
VOCs detection for field applications. The adaptive sampler is based on
the use of a front-end solid phase microextraction (SPME) array that
passively extracts volatiles from large air volumes. The sampler is
coupled to a microconcentrator for overall flow matching and is interfaced to a low thermal mass (LTM) gas chromatograph (GC) employing
both mass spectrometric (MS) and pulsed Helium ionization detection.
SPME was selected as a passive air extractor because the technique
requires neither power nor solvents for sample collection or preparation,
and the array device can be made compact. Trapping, desorption, and
subsequent microconcentration efficiencies will be presented for quantitative VOC studies. The theoretical aspects of SPME indicate that in all
but the smallest of samples, sample size (i.e., volume of water or air)
does not affect the sample loading onto the fiber. However, analyte concentration and sampling conditions do affect quantitative fiber loading.
Currently GC/MS is used in the laboratory to confirm the identity of a
standards mix as well as a spiked scent pad; however, data from this new
small, lightweight, and low power prototype sampler will be used to
improve GC/MS field systems.
The laboratory division and forensic community should benefit
from these method and instrumentation developments because it will
help reduce the time for mixture analysis of the various organic matrices,
including semivolatiles as well as volatiles, typically encountered by
responders and analysts in the field.
VOCs, Scent, Adaptive Sampler
C43
Certified, My (Beep)!
Denise A. Shepperd, BS*, Trillium, Inc., 2014 Carol Drive, Wilmington,
DE 19808; Carol A. Erikson, BS, MSPH, Trillium, Inc., 356 Farragut
Crossing Drive, Knoxville, TN 37922
certification of a laboratory by some agency does not guarantee that the
results they produce will be valid.
humanity by demonstrating that analytical results can be wrong even
when they are reported by a “certified” laboratory.
Why on earth would anyone want to validate pH data? It’s a no136
brainer, right? You get the meter ready, standardize with a couple of
buffers and start analyzing. This is not rocket science.
Well, let’s just pretend that you are asked to validate, for some
unknown reason, the pH data for a set of samples. And say those samples
happen to be soils. Everything looks good in the raw data, the standards
look good, replicate readings are close, and the de-ionized water gives
consistent readings. Not much to say. It looks like the results are good.
But wait, the results on the summary forms in the data package don’t
agree with the results on the final reports. As a matter of fact they differ
by two, three, even four pH units. Not only that, but the laboratory also
gave sample-specific reporting limits for pH and they vary from soil
sample to soil sample. What happened? You won’t believe the answer to
that question. Maybe validating that data wasn’t such a bad idea after all.
Next, we’re looking at data for volatile organic compounds by EPA
Method 524. The laboratory purges 25 mLs of a water sample and analyzes it with a benchtop mass selective detector (MSD). The calibration
curve is established between 0.5 ug/L and 50 ug/L. That means, from the
instrument’s point of view, the calibration represents a range of 12.5
nanograms to 250 nanograms of the target compound in the column.
That’s a reasonable range for most compounds on an MSD.
But what is the lab reporting? In this particular instance, the laboratory reported carbon disulfide in one water sample at 0.041 ug/L and
1,1-dichloroethane at 0.077 ug/L in another. Both results were qualified
as estimated, as they well should be, but what about these numbers?
According to the calibration, they can’t accurately quantify anything
below 0.5 ug/L. So why are they reporting a result down to 0.041 ug/L?
Another good question, with a not-so-good answer.
The laboratories that reported the results described above happen to
be “certified.” But what , exactly, does certification mean? When you
are remediating a site, reporting data to an agency, or using data to
support some other action on a project or site you will often hear, “we
used a certified lab,” or “we want to make sure the lab is certified,” or
“you need to use a certified lab, we’ll only accept results from a certified
lab.” Okay, so you used a certified lab, but does that mean you’re going
to be getting good results? Maybe. Maybe not.
The incidences described above, as well as a number of other
common and uncommon laboratory mistakes gleaned from experiences
in data validation, will be presented to illustrate that results can be
wrong, even from a certified laboratory.
Certification, pH Analysis, Calibration
C44
Forensic Investigation of an
Underground Gas Main Explosion
Chin-Chin Lim, MSc, MBA*, Poh Ling Chia, BSc, and Ming Kiong
Michael Tay, PhD, MBA*, Centre for Forensic Science, Health
Sciences Authority, 11 Outram Road, Singapore 169078, Singapore
The goal of this presentation is to present a case study of a forensic
investigation into the explosion of a buried section of a town gas
distribution system.
humanity by illustrating how forensic scientists can complement
structural design and civil engineers, as well as gas safety experts, in
determining the cause of engineering failures.
Background
The explosion occurred in a single-story seafood restaurant erected
on a 35-cm thick reinforced concrete (RC) floor slab on land reclaimed
from the sea 30 years ago. This concrete base had two layers of reinforcement bars (re-bars) and was supported by large pilings driven into
the ground. As a result of soil settlement, there was a void of 30-50 cm
under the monolithic concrete slab, apparently extending throughout the
entire space under the restaurant. The rear area of the restaurant was a
fairly open concrete structure with corrugated iron roofing, false ceiling
and thin-walled partitions.
* Presenting Author
The underground town gas main entered the restaurant at the rear
area. The piping consisted of two levels of 4-inch gas pipes joined
together by an inclined spigot. At the upper level, the inclined spigot was
connected by a spigot-and-hub joint to a horizontal pipe, both of which
were embedded in a concrete channel in the RC slab. The town gas in
the main contained hydrogen (62%), carbon dioxide (18%), carbon
monoxide (6%), methane (5%), pentane (5%) and traces of nitrogen,
oxygen and a malodorous mercaptan. Although this pipeline was still
charged, it had been capped for three years. The restaurant had converted
to using LPG cylinders in a manifold in the kitchen and dining area.
In response to a major leak in the underground water main, located
in the open dining area, two workmen opened the water valve pit, ventilated it for an hour and began working around the leaking water valve.
About 30 minutes later, as they were hacking tiles in the pit with an iron
spike and an electrical drill, a series of explosions occurred, The first
explosion occurred in the valve pit, followed by a series of directional
explosions moving underground towards the rear of the restaurant where
the town gas main was located. The explosions covered the premises
with debris, smoke and dust but no fire resulted. The injured workman
recalled smelling cooking gas immediately after the first explosion.
The explosions damaged the ceiling, roofing and walls, popped
floor tiles, and dislodged furnishings. The rear area where the underground town gas main entered the premises sustained the most severe
damage. The explosion broke the concrete channel in which the gas pipe
was embedded, exposing the 45o elbow (bend) attached to a horizontal
pipe, and a void underneath with a disconnected inclined spigot partially
buried in subsided soil. The horizontal pipe attached to the elbow was
partially resting on the lower layer of re-bars. The upper layer of re-bars
had been cut and removed to create space for embedding the horizontal
section of pipe during installation.
The explosion also dislodged the LPG cylinders which were all
intact and clearly not the cause of the explosion. Except for two blown
out sewage inspection chamber (IC) manhole covers in the open eating
area, all of the manhole covers were intact, and waste-water in these ICs
was found to be free-flowing. Property management personnel indicated
there was no foul smell coming from the underground sewage system
before the explosion.
The town gas company promptly shut off the gas supply to the area
after the explosion. Two structural engineering consultants hired by the
gas company to investigate the incident independently concluded that
the separation of the 45o elbow joint was sudden, was unlikely to have
been caused by natural soil settlement, and was a result rather than the
cause of the explosion. One consultant dismissed the possibility that the
explosion was due to leakage of town gas.
Key issues
• Our lab was tasked to verify the consultants’ findings and examine
the following issues:
• Was the dislodged joint caused gradually and progressively by the
settlement of soil? Or was the joint failure sudden and catastrophic?
• Was the explosion caused by methane produced either by the
reclamation soil in-fill or underground sewage system?
• Was the explosion caused by town gas leaking from the gas
pipeline?
Analysis of residual gases
Holes were drilled at 5 locations through concrete floor and the
void underneath these holes was analyzed for hydrogen and methane but
none was detected. Less than 50 ppm methane (well below the 50,000
ppm LEL for methane) was found in the IC manholes. Soil samples collected from around the damaged gas pipe, and from the water valve pit,
were examined for methane, ethane, propane and butane, but none was
detected.
Examination of the dislodged joint
The inclined spigot and the 45o elbow attached to the horizontal
pipe were cut and brought to the lab for further examination. The cut-out
spigot was not deformed or dented. However, its inner surface and end
(circular ring) were rusty.
* Presenting Author
The 45o elbow (bend) had two hubs, one of which was attached to
the horizontal pipe. A flexible optical fibre scope revealed rust spots and
patches on the interior surfaces of both the hub and the pipe, indicating
the presence of moisture and oxygen within the piping system for some
time. In addition to rust spots, the interior walls of the hub had a circular
arc of rust corresponding to the end of the spigot. This arc of rust, evidently formed by prolonged contact with the end of the spigot, was not
concentric, indicating that the spigot was not aligned axially to the hub.
The joint was mechanical with a seal effected by compressing a
wedge-shaped annular gasket of elastomeric material, partially lined
with lead sheath, onto the jointing surface in the pipe hub and the outside
of the pipe spigot, by means of a pressure gland and a series of bolts and
nuts. This joint was carefully dismantled and examined. Rust and fine
soil particles were found on the tapered inner surface of the hub where
the gasket had been in close contact with the iron surface. The gasket had
suffered extensive wear and tear. The elastomeric material was permanently deformed and flattened. The lead sheath was torn and severely
thinned out on one side, and stained with rust or brownish soil stains.
The lead layer had buckled furrows with dirt and fine soil between the
lead and elastomeric layers, indicating that the seal was not gas-tight.
The furrows evidently resulted from the spigot end pressing against the
top part of the hub, creating a disproportionate amount of pressure along
the top of the annular gasket, which thinned and extended out the lead
along its circumference, creating folds in the wider available space. The
inner surface of the gasket indicated uneven contact with the spigot end
and the gasket was not evenly seated around the entire joint.
Findings
The most probable cause for the explosion was a damaged gasket
resulting from a badly aligned and severely stressed joint. This faulty
gasket was not gas-tight; the leak led to a diffusion and accumulation of
town gas in the connecting spaces between the monolithic concrete slab
and the subsided soil. The electric drill ignited the flammable gas and
triggered the explosion, which generated a significant expanding force in
the underground space, lifting up and breaking the weak concrete floor
channel bearing the horizontal pipe. The uplifting raised the horizontal
pipe and attached elbow, and created a sudden longitudinal pullout force,
which caused a sudden axial withdrawal and separation of the 45o elbow
joint.
The misalignment of the joint was probably due to the settlement of
soil under the buried inclined spigot, which drastically reduced support
for this underground segment. Severe stress was created at the elbow
joint because the embedded horizontal pipe and the elbow were firmly
supported by the lower layer of re-bars in the concrete channel, whereas
the inclined spigot pipe being inadequately supported, was sinking. The
weight of this partially buried spigot in subsiding soil created a concentration of stress on the joint.
Conclusion
Our investigations revealed departures from industry practices concerning installation of the joint and support of the piping system. Failure
to account for soil settlement and provide adequate long-term support
for the inclined spigot and lower level of pipes resulted in misalignment
and sagging. Although gas pipelines are commonly designed for 50
years of service, the forced dislocation (deflection) of the piping and fittings eventually took the joint to its limit, creating leakage paths between
the spigot pipe and the gasket. A continuous outflow of town gas ensued
as the pipe was pressurized.
Detailed examination of the joint was crucial in this case, over and
above general observations about damages and speculations over different possible contributing factors. This case illustrates how forensic
scientists can complement structural design and civil engineers, as well
as gas safety experts, in determining the cause of engineering failures.
Gas Pipe, Stressed Joint, Gasket Failure
137
C45
Virtual Loosening of Fittings
From Elevated Temperatures
Derek T. Nolen, BS, MSME* and Jean L. McDowell, BSc, McDowell
Owens Engineering, Inc., 1075 Kingwood Drive, Suite 100, Kingwood,
TX 77339; E. Philip Dahlberg, PhD, Metallurgical Consultants, 7701
Parnell, Houston, TX 77021; Kevin Tajkowski, MS, McDowell Owens
Engineering, Inc., 1075 Kingwood Drive, Kingwood, TX 77339
After attending this presentation, attendees will have a basic understanding of why threaded fittings may be loose after being subjected to
elevated temperatures, such as from exposure to a fire.
humanity by demonstrating the importance of applying engineering
sciences in fire investigations to correctly analyze and interpret the
evidence.
A fire occurs at a property supplied by natural or LP gas. Following
the fire, investigators test the gas system, find leaks, and ultimately
implicate the gas company or pipe installer for faulty workmanship.
Was the connection indeed loose prior to the fire? Barring unforeseen
circumstances (tampering), the fitting was likely leak-free at the time the
fire started, particularly if proper leak tests were performed at the time
of installation. The naïve investigator reasons that the only way for a
fitting to be loose is for it to have been loose prior to the fire, but it is not
the torque that should be of sole interest after the fire, rather the number
of threads engaged. The number of threads engaged becomes the only
reliable method of estimating the pre-fire engagement torque.
Looseness in pipe and tubing fittings post-fire must be carefully
scrutinized before an assumption of pre-existing leaks can be made.
Investigators must understand that sufficient heating of threaded metal
fittings, as in basic heat treatment processes, relieves stress in the metal.
Stress in the metal is what allows a fitting to become tight and “leak
free.” Heat from a fire, if not sufficient to completely melt the fitting,
can allow the metal to re-crystallize, thus softening and relieving stress
in the fitting. This results in a virtual loosening of the fitting, although
no rotation or unscrewing of the parts has occurred.
Experiments conducted with various flare and pipe fittings of different materials showed that brass flare fittings, a form of compression
fitting, “loosened” to the greatest degree with the lowest temperature
exposures. Steel pipe fittings required the greatest heat exposure and
resulted in the least effective “loosening.” “Leak free” fittings with
known, standardized initial torques were subjected to elevated temperatures for various times. After removal from the elevated temperatures,
the fittings were tested for leaks and post heating torque values. These
results demonstrate how stress-relieving processes can, and do, occur in
environments like house fires. The simple engineering process of stress
relief and how it relates to threaded fitting tightness must be understood
by investigators involved with testing gas systems following fires in
order to properly interpret the fitting conditions, and how they relate to
pre-fire conditions.
Loose, Fittings, Fire
C46
Bad Science From Big Brother
Beryl Gamse, PhD*, Jean L. McDowell, BSc, Derek T. Nolen, MS, and
Nestor Camara, McDowell Owens Engineering, Inc., 1075 Kingwood
Drive, Suite 100, Kingwood, TX 77339
After attending this presentation, attendees will have an appreciation for the need to critically evaluate published research, particularly
that done by authoritative agencies.
humanity by illustrating the importance of preventing unreliable reports
from being used as the basis of opinions in litigation.
It is essential that we, as scientists and engineers, do good science.
It is especially important when this science is the basis of our opinions
138
that we present to lay people in the course of litigation. It is also
incumbent upon us to be vigilant in detecting bad science that may
become current and the basis of opinions in the future, particularly if the
bad science carries the imprimatur of a federal regulatory agency.
As part of a project to develop standards addressing the potential
hazards of clothes dryer fires, the Consumer Product Safety Commission
conducted a program that resulted in the publication, “Final Report on
Electric Clothes Dryers and Lint Ignition Characteristics,” May 2003. In
the test program, a great deal of effort was put into establishing some of
the operating characteristics of several electric clothes dryers and
demonstrating certain conditions under which lint, taken from a clothes
dryer lint screen, will burn when positioned on or near an energized
clothes dryer heater unit that had been removed from a dryer and placed
in a special test chamber. Very little of the report was devoted to lint
accumulation, apparently because very little accumulated during their
testing. The report indicates that: 1) lint collects on the heater unit
housing in locations and at densities similar to the samples tested, 2)
large clumps of lint can become located within a few inches of the heater
unit entrance along its longitudinal axis , and 3) the lint so located could
ignite and create larger fires in the cabinet or downstream of the heater
unit in the clothes-filled tumbler drum, at the lint screen, and/or in the
exhaust vent. There was nothing in the report that established that lint,
similar in density and quantity to that used in the lint ignition tests, could
become positioned relative to the heater unit as it was in the tests. In
fact, our experience in laboratory testing under realistic operating conditions and examining dryers after years of service in homes contradict the
first two items. Thus, the report gives a false impression relative to the
hazard of dryer fires.
In addition to not establishing the lint accumulation, the test conditions for ignition of that accumulated lint were not valid.
The test chamber for lint ignition tests was essentially a horizontal
wind tunnel with the dryer heater unit mounted with its longitudinal axis
and internal air flow vector horizontal and its sides isolated from the test
chamber walls. In most dryers the unit is oriented vertically and one of
its sides is in contact with the metal tumbler support bulkhead. Neither
the flow field nor the heater unit surface temperature distribution were
documented in the actual dryer, therefore the flow fields and distributions created in the test chamber could not be validated.
Our testing has shown that, in operating dryers of the type tested by
CPSC, the flow is downward in the cabinet, opposite to that used in their
ignition tests. The downward flow continues past the heater inlet with
only a portion of air being drawn into the heater. The general downward
flow prevents any lint that is more than a few inches below the inlet from
possibly being drawn into the heater unit. Thus, no lint accumulated on
the bottom of the cabinet can be drawn to the heater. Also, the orientation
of the gravity vector in the test chamber is incorrect. A clump of lint that
is somehow located in the limited in-draft at the entrance to the heater
unit may be too dense to be drawn into the unit if the unit is oriented vertically but not if it is located horizontally. These factors make their
testing and conclusions relative to lint ingestion problematic, if not
invalid.
The configuration used for the ignition of materials downstream of
the heater has no relationship to an actual dryer. The target material, lint
or cotton toweling, was placed immediately downstream of the horizontal heater unit in a 4-inch diameter duct. The air flow speed across
the target material, which restricted the cross-section of the duct, was not
reported, but was probably significantly higher than the 800 sfpm at the
heater inlet. This air flow would, and did, fan any smoldering embers
into flames. In an actual dryer, the exhaust of the heater passes through
a perforated metal wall before reaching toweling-type materials, then
traverses the tumbler plenum and, in some designs, another perforated
metal wall to reach lint on the collection screen, and then pass through a
centrifugal blower to reach any lint deposits in the exhaust vent. The perforated metal walls act as spark arresters eliminating most, if not all, of
the lint embers. The directed airflow goes to essentially zero in the
plenum chamber of the drum. The only air movement would be due to
* Presenting Author
the tumbling clothes, which would probably beat any surviving embers
out of existence. The survival rate for lint embers reaching the lint screen
or beyond would be negligible. No explanation is given as to how lumps
of lint could make their way into the heater unit. Even if one accepts that
somehow clumps of lint are ingested into the heater unit, it must be
demonstrated in an actual dryer that embers from it will survive in sufficient size and quantity to cause ignition of material downstream of the
heater unit.
The report covers a large number of tests which are described in
great detail; many data are presented which are analyzed and discussed
extensively. The extent of the discussion and the fairly attractive presentation of the voluminous data divert the readers’ attention from the fact
that, though what is presented follows the two subjects mentioned in the
title of the report, “Electric Clothes Dryers” and “Lint Ignition
Characteristics,” no valid connection is made between the two. The
testing program and report are examples of bad science.
Dryers, Flint Fires, CPA Safe
C47
Variation in Performance of Residential
Smoke Detectors — Safety Concerns
Due to Unpredictable Behavior
B. Don Russell, PhD* and Carl Benner, MS, Texas A&M University,
238 Wisenbaker, MS 3128, College Station, TX 77843-3128
The goal of this presentation is to educate forensic engineers as to
the wide variation in performance of commercial residential smoke
detectors under similar fire conditions as an aid to forensic investigation.
humanity by describing how the performance evaluation of smoke
detectors will help educate forensic scientists and engineers who are
called upon to perform first fire analysis of detector performance and circumstances of injury to fire victims. The performance data will also
assist fire safety professionals in determining ways to improve the performance of smoke detectors, thereby increasing public safety and
reducing the loss of life and property resulting from fire.
This paper presents the results of an extensive research project
evaluating residential smoke detectors under controlled fire tests. The
performance of residential smoke detectors has been extensively documented using full scale testing. Fire scenarios include those typical and
expected in residential environments under circumstances that pose
hazard of injury or death to residents.
Commercially available, UL-listed smoke detector designs from
multiple manufacturers have been tested in large quantities under a wide
variety of fire scenarios. Specific emphasis has been given to the performance of same make/same model detectors under similar fire circumstances. The objective is to determine the statistical predictability of
the performance of detectors as purchased, installed, and used by the
public, as a function of various fire scenarios common to residences in
the United States. The results are presented blind, without specific reference to manufacturer, and without identification of a specific product.
The test protocol utilized requires testing of unmodified, unaltered
smoke detectors purchased commercially. Detectors were purchased in
multiple units of the same make/same model for comparison purposes,
but no attempt was made to control for date of manufacture or shelf time
prior to sale. Smoke detectors were installed in a full-scale residence
utilizing the manufacturer’s recommended installation instructions.
Fires were set in the residence consistent with foreseeable fire scenarios
in private housing in the United States. A common fire scenario to which
detectors were tested is smoldering/burning furniture or bedding.
The full-scale residence was instrumented to provide detailed parametric information documenting the progression of the fire. Computer
based instrumentation records such parameters as temperature, obscuration in multiple locations, and the performance of individual detectors
* Presenting Author
in response to smoke production. For each detector, performance criteria were recorded and documented to include the time each detector
takes to respond to the fire and the obscuration levels in and around the
detector and in the residence at the time each smoke detector responds.
From these data, relative comparisons were made between same
make/same model detectors and different make/different model
detectors in all combinations. These data were then used to draw conclusions with respect to overall performance, efficacy of the detectors
for providing home safety, and the predictability of smoke detector performance based on fire type.
The intent of this paper is to educate forensic investigators and
forensic engineers concerning the actual performance of smoke
detectors in residential fires as compared to the performance that is
expected by the public or by engineers and fire scientists. The results of
research to date show wide variation in performance by units of the same
model or essentially same model of detector, making prediction of
behavior difficult in post fire analysis. The characteristics and factors
affecting performance and variations in performance are described and
presented. Specific case studies are reviewed with research data from
actual fire tests presented and discussed. Overall performance of
detectors as a function of tenability parameters, such as obscuration, is
presented.
Smoke Detectors, Fire Detection, Fire Safety
C48
Analyzing a Starter-Generator Failure
on a Turbo-Prop Aircraft Engine
Raymond K. Hart, PhD, JD*, 145 Grogan’s Lake Drive, Atlanta,
GA 30350-3115
The goal of this presentation is to describe a unique item of electrical equipment which performs two separate tasks in the operation of
small turbo-prop and turbo-jet aircraft engines. While in flight, function
lights indicated a problem had occurred with the electrical generator on
one of the aircraft’s engines, so the airplane made an emergency landing.
Instead of waiting for a certified FAA mechanic to inspect the engine the
next morning, a cursory inspection was made by the pilot, and finding
nothing obviously wrong, he attempted to restart the engine and the
result was disastrous.
humanity by demonstrating why an operator of any equipment
should always rely on the vehicle’s instruments, and not try to
out-smart those instruments.
There are thousands of one and two turbo-prop engined aircraft
flying today. Should a pilot, having seen a generator malfunction light
illuminate during flight and tried to trouble-shoot the problem, it could
have roved to be disastrous to life and aircraft. Manufacturing defects
have largely been engineered out of present day Starter-Generators and
thus contributing to greater safety for passengers and crew while flying
small jet-powered aircraft.
A chartered Piper Cheyenne aircraft was returning to Sarasota, FL,
from Miami, FL, when the pilot noticed a generator red “NO-CHARGE”
light illuminate. He immediately changed course and landed at Naples,
FL, airport. He removed the left engine covers and visually inspected
the auxiliary equipment in the left side PT6A-41 engine, and in particular, the AUXILEC Starter-Generator. The pilot determined the
brushes were not worn away and that they were functioning correctly,
and after taping other relays in the generator circuit he decided to restart
the engine. After a second or two, and with the generator light still on,
the starting mechanism completely seized.
The subject Starter-Generator, DC Aircraft, was an AUXILEC
Model 8013C. Its operating specifications as a starter were 36 volt max.
at 1000 amp max., and as a generator its output was 30 volt at 200 amp,
with a shaft speed of 7000/12150 rpm.
139
From the air-intake end of the PT6A-41 engine, which faces
rearward, the compressor turbine was connected to the outside of that
gear box. The two shafts were linked together through bevel gears which
had an approximate 3:1 gear ratio. During the engine starting procedure,
the starter section of the unit is powered from the battery and it runs-up
the compressor to approximately 30,000 rpm and then the engine’s operation is commenced by the introduction of fuel to the combustion
chamber and energizing the ignitors. A generator reverse current relay
senses a reduced current requirement in the armature series field coils,
and that relay switches to the output from the shunt-field armature coil
in the then driven Starter-Generator.
The illumination of the generator charge light during flight was
indicative of an electrical fault, which can often be traced to heat damage
to insulation on electrical components, and lead to electrical shorts, or
electrical/mechanical malfunction between the brushes and the commutator. After an emergency landing the pilot removed the engine covers,
and with a flashlight and screw driver he proceeded to inspect the then
exposed commutator end of the starter-generator where most electrical
faults have historically been found. In the subject case nothing was
found to be out of order, so the pilot then attempted to restart the engine.
After several seconds of applying battery power to the starter, the
armature abruptly stopped turning, and it was only then that the pilot
realized the futility of trying to restart the engine, and so he left it to be
inspected by a mechanic the following morning.
A partial tear-down found the bevel drive gears in the assessory
gear box had been electrical-resistance welded together. Further, four
(4) lengths of the forward facing beryllium-copper (Be-Cu) armature
retaining ring were recovered from the inside of the front housing. The
armature series-field coil windings and the shunt-field coil windings had
been rubbed, or machined, to expose bare copper wire.
Rubbing-wear signatures were observed on one portion of the
armature’s circumferential surface as well as on one portion of the facing
surface in the stator. Those rub marks indicated the armature had rotated
in an elliptical pattern for a considerable period of in-service time. The
temperature excursions due to frictional rubbing of the metal components in the armature and stator were accompanied by thermal
expansion, heat treatment and oxidation/corrosion of the metal parts. At
a late stage in the stator-generator service life the armature ring cracked
all the way across its width, starting at one of the incipient cracks in the
surface of the hydrogen-embrittled Be-Cu ring - as determined by
SEM/EDS analysis.
The longest piece of the Be-Cu ring had been bent into a “J” shape
with the hook-end around the shaft and the free end long enough to make
contact with the bare copper ends of the armature coils. The shunt-field
coil had been partly shorted to the armature shaft by the “J” piece,
causing the generator light to illuminate. During normal engine operation, the starter series-field coils are disconnected from the battery, so
no serious damage was done when the bare copper wire ends of the
starter coils were also shorted by the metal “J” piece.
When an attempt to restart the turbine was made, the full charge of
the battery was shorted across the metal “J” piece to the armature shaft.
However, the path of least resistance to the “engine ground” was through
the bevel gears in the auxiliary gear box. They became welded together,
and their seizure caused considerable damage and expensive repairs to
both the accessory gear box and to the compressor turbine.
The case went to trial, and the verdict was that the starter-generator
failed due to a manufacturing defect, but all other repair costs were the
responsibility of the pilot/owner, who should not have attempted to
restart the engine.
Turbo-Prop Engine, Starter-Generator System, Beryllium-Copper
Armature Ring
140
C49
The Characterization of Binary-Output
Walkway-Safety Tribometric Instruments
by Characteristic Functions
Mark I. Marpet, PhD, PE, St. John’s University, 300 Howard Avenue,
Staten Island, NY 10301; Howard P. Medoff, PhD, PE*, Pennsylvania
State University, Woodland Road, Abington, PA 19001
After attending this presentation, attendees will learn about using
logistic regression parameters to characterize the response of binaryoutput tribometers. This information can be used to determine if different tribometers give equivalent results.
humanity by demonstrating a method of characterizing walkway safety
tribmeters that give binary, i.e., slip/no-slip results. The characterization
gives insight into the accuracy and repeatability of the tribometer.
One way of broadly characterizing tribometers is to classify them
as to whether they give a quantitative or a binary output. A drag sled, for
example, gives a quantitative output (the measurement of the relative
lateral force is an indicator of the friction). The Portable Inclinable
Articulated Strut Tribometer (PIAST) is an example of a tribometer that
gives an output in binary form. The carriage is set and locked at a given
slip angle and the test foot is released. Either the test foot slips or it does
not slip (and hence, the output is binary). Ideally, the tribometer will slip
100% of the time at available-friction values below the ‘set’ friction
value and never slip at available-friction values at or above the set
friction. (The ‘actual friction’ is a function of the friction measuring
system, so comparisons of characteristic curves generated by different
types of tribometer should be approached with caution.)
As with any idealized phenomenon, what we observe in fact is only
an approximation to this idealization. What actually occurs is that there
is no sharp-cornered vertical line delineating the slip/no-slip boundary.
Depending upon the tribometer and the test conditions, the actual
boundary has both a slant and rounded corners; at any given friction
value near the actual-friction value, there is a finite probability, p, that
the tribometer test foot will slip and a complementary probability, 1-p,
that it will not.
A statistical tool that can characterize such situations is Logistical
Regression (LR). LR uses the occurrence or non-occurrence of a qualitative variable (here slip or no-slip) as the dependant variable and a set
of continuous parameters as the independent variable set. (Here, the
friction of the tribometric system is the single independent variable.)
Essentially, LR uses a least squares, maximum likelihood approach with
the negative sum of the logs of the probabilities attributed to the
response levels that actually occurred at each observation. (Note that,
because all probabilities are between 0 and 1, all the logs are negative.)
The regression parameters can be used to characterize the tribometric
system.
* Presenting Author
C50
Walkway Slip-Resistance Tribometry
With Elastomeric Test Feet
Robert H. Smith, PhD, PE*, National Forensic Engineers, Inc., PO
Box 82486, Kenmore, WA 98028
The experimental set-up is as follows (all stationary parts except
the platform rail have been omitted for clarity). A level platform is constrained to move in a rectilinear fashion by low-friction instrument
bearings bearing against a platform-support rail. To this platform,
attached by string and pulleys, is a counterbalanced weight platform.
When the platform is weighted to simulate a lateral force, it rests against
a stop; the tribometer exerts a lateral force in a direction opposite that of
the force exerted upon the platform by the weight. When the tribometer
is released, the test foot contacts the platform, which is covered with
double-stick tape, so as to eliminate all lateral force except as provided
by the weight and the dynamic effects of the tribometer. The tribometer
is set at a given level and repeatedly released. The dependant variable is
recorded for each trial, i.e., slip/no-slip. The tribometer setting is varied
and the process repeated. The weight is varied from the point where the
carriage slips each and every time to the point at which the carriage
never slips. The resulting data is analyzed using logistic regression.
(Vertical load = 5.23 kg.)
Forensic Engineering, Walkway-Safety Tribometer, Slip-and-Fall
Accidents
* Presenting Author
The goal of this presentation is to inform the forensic safety community that situations exist in which the classical coefficient of friction
equation can be inappropriate for use in quantifying walkway traction
characteristics.
humanity by demonstrating application of a methodology which will
allow calculation of the adhesion component of elastomeric friction on a
scientific basis.
In walkway slip-resistance metrology a constant coefficient of
friction equation is utilized as both the empirical and rational relationship
to quantify the traction characteristics of surfaces encountered during
walking. However, rubber is widely employed in footwear. In 2001,
various types accounted for 55.3 percent of worldwide shoe heel and sole
material usage. It has been shown repeatedly that rubber exhibits a
decreasing coefficient of friction with increasing applied load. Many
pedestrians experience this situation in everyday ambulation. When such
occurs, the friction coefficient will not only decrease, but vary with pedestrian weight. It has also been shown that decreasing friction coefficients
can be produced in the testing regimes of certain static and dynamic tribometers utilizing elastomeric test feet. Although use of constant coefficients to quantify walkway skip resistance in comparison to the generally
accepted standard of 0.5 has been employed for many years, they have
been justified only on an empirical, experiential basis. Use of constant
elastomeric coefficients in such circumstances has not been scientifically
justified. A comprehensive foundation for application of static and
dynamic friction theory applicable to the elastomeric materials involved in
walkway slip-resistance metrology has not been presented.
A review of the literature revealed that constant coefficients of
friction have been utilized in walkway slip-resistance metrology since
about 1930. At that time, Hunter designed his articulated strut tester to
measure the angle at which a leather test foot slipped on selected walkway
materials when an 80-lb weight was applied. The tangent of the slip angle
was taken as the coefficient of friction. The constant coefficient of friction
equation (Amontons’ law) was originally developed in classical metallic
theory in which the static and dynamic coefficients equal the tangent force
resisting slippage of smooth, contacting metal surfaces divided by the
normal load. Application of constant coefficients in the assessment of
walkway safety presumably arose from their wide application in metallic,
machinery operation where minimization of metal-to-metal contact resistance is desired. In such circumstances, contacting asperities on the metal
surfaces are predominantly in the plastic range. The significant friction
forces developed between these asperities comprise atom-to-atom
adhesion. This mechanism is characterized as cold welding. When a shoe
heel is an elastomer, its deformation during ambulation will likely remain
elastic. Furthermore, because of their elasticity, elastomers experience
both atom-to-atom and van der Waals adhesion.
Considerable investigation has been carried out on frictional characteristics of elastomers employed in fields other than ambulation safety.
Findings from these efforts have applicability in walkway slip-resistance
tribometry. An empirical and rational relationship, derived from the
Hertz equation, has been developed for quantifying friction force generation when coefficients of friction decrease. The physical mechanism at
work in these circumstances involves real areas of contact, or near
contact, of elastomeric asperities and a smooth walking surface, where
either type of adhesion develops. When applied normal loads increase,
the areas of contact, or near contact, also increase. As a result, the
friction force increases. However, its rate of increase can be less than
that of the applied load and their ratio falls. Thus, a decreasing coefficient in these conditions does not indicate decreasing frictional resistance.
141
When applied to elastomeric materials, the Hertz equation takes the
form FT = c(FN)m; where FT is the developed, tangent, friction force, c
is a constant associated with the Young’s moduli and Poisson’s ratios of
the two materials involved, FN is the applied normal load, and 2/3(m(1.
It must be emphasized, however, that the Hertz equation quantifies only
the adhesional component of friction. Other friction force mechanisms
exist when elastomers slide on a harder surface. The Hertz equation
should be applied only after the frictional resistance arising from the
other mechanisms is identified, quantified, and subtracted from the total,
measured tangent force. This capability awaits development of a reasonably complete understanding of elastomeric friction as it applies to
pedestrian ambulation.
Variable Coefficients of Friction, Elastomeric Shoe Heels,
Walkway Slip Resistance
C51
Human Posture Control: Preparation
Gait to Avoid Slips and Falls
Woo-Hyung Park* and Simon M. Hsiang*, T

Volume 10 - American Academy of Forensic Sciences

Transcription

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